1、raising standards worldwideNO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBSI Standards PublicationBS EN 16342:2013Cosmetics Analysisof cosmetic products Quantitative determinationof zinc pyrithione, piroctoneolamine and climbazole insurfactant containing cosmeticanti-dandruff
2、 productsBS EN 16342:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 16342:2013.The UK participation in its preparation was entrusted to TechnicalCommittee CW/217, Cosmetics.A list of organizations represented on this committee can beobtained on request to
3、its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2013. Published by BSI StandardsLimited 2013ISBN 978 0 580 76542 1ICS 71.100.70Compliance with a British Standard
4、cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 May 2013.Amendments issued since publicationDate Text affectedBS EN 16342:2013EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16342 May 2013 I
5、CS 71.100.70 English Version Cosmetics - Analysis of cosmetic products - Quantitative determination of zinc pyrithione, piroctone olamine and climbazole in surfactant containing cosmetic anti-dandruff products Cosmtiques - Analyse des produits cosmtiques - Dtermination quantitative de la pyrithione
6、de zinc, de la piroctone olamine et du climbazole dans les produits cosmtiques antipelliculaires contenant des agents de surface Kosmetische Mittel - Untersuchung von kosmetischen Mitteln - Quantitative Bestimmung von Zinkpyrithion, Pirocton-Olamin und Climbazol in tensidhaltigen kosmetischen Mittel
7、n mit Antischuppenwirkstoffen This European Standard was approved 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 l
8、ists 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 und
9、er 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
10、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 EUR
11、OPEN 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 16342:2013: EBS EN 16342:2013EN 16342:2013 (E) 2 Contents Page Foreword
12、3 Introduction .4 1 Scope 5 2 Terms and definitions .5 3 Principle 5 4 Reagents .5 5 Apparatus and equipment 7 6 Sampling .7 7 Procedure .8 8 Evaluations .9 9 Test report . 10 Annex A (informative) Results of the inter-laboratory test . 11 Annex B (informative) Sample Chromatogram 12 Bibliography .
13、13 BS EN 16342:2013EN 16342:2013 (E) 3 Foreword This document (EN 16342:2013) 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 standard, either by publication of an identical tex
14、t 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 possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held respon
15、sible 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, Finlan
16、d, 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. BS EN 16342:2013EN 16342:2013 (E)
17、4 Introduction Special hair products contain substances to help prevent dandruff. These substances mainly inhibit the development of microorganisms, which often are the cause of dandruff. The most commonly used substances are zinc pyrithione, piroctone olamine and climbazole. The substances are regu
18、lated by Council Directive of 27 July 1976 on the approximation of the laws of the member states relating to cosmetic products (EC 76/768/EEC) as well as Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products. Limits for these substances a
19、re listed in the annexes regulating preservatives in cosmetic products. Zinc pyrithione is additionally listed in Annex III of both regulative documents named above. NOTE As the Regulation (EC) 1223/2009 applies in total from 11 July 2013 and replaces Directive 76/768/EEC the following details relat
20、e only to Regulation (EC) 1223/2009. Reference Number, maximum authorised concentration in hair products, limitations and requirements: Annex III Regulation (EC) 1223/2009 Zinc pyrithione: No. 101: 0,1 % leave-on hair products Remark: For purposes other than inhibiting the development of microorgani
21、sms in the product. This purpose has to be apparent from the presentation of the product. Annex V Regulation (EC) 1223/2009 Zinc pyrithione: No. 8: 1,0 % hair products Remark: Only in rinse-off products 0,5 % other products Remark: Not to be used in oral products Climbazole: No. 32: 0,5 % Piroctone
22、olamine: No. 35: 1,0 % rinse-off products 0,5 % other products BS EN 16342:2013EN 16342:2013 (E) 5 1 Scope This European Standard specifies an analytical method for the detection and quantitative determination of the following anti-dandruff agents: zinc pyrithione, piroctone olamine and climbazole i
23、n surfactant-containing cosmetic products in the concentration range from 0,1 g/100 g to 1,0 g/100 g. NOTE The method is also suitable for the determination of ketoconazole and ciclopirox olamine (q.v. Annex A) in surfactant-containing cosmetic products and it is probably applicable for the determin
24、ation of the substances in non surfactant-containing cosmetic products. For these purposes, the method has not been validated. 2 Terms and definitions For the purposes of this document, the following term and definition applies. 2.1 anti-dandruff agents substances, added to hair care products, activ
25、e against the development of microorganism e.g. zinc pyrithione, piroctone olamine and climbazole 3 Principle The anti-dandruff agents are extracted from the cosmetic sample matrix using dichloromethane and methanol. Each analyte present in the sample extract is separated using reversed phase HPLC w
26、ith UV (DAD) detection. The quantitative determination is made using the external standard method of calibration. 4 Reagents 4.1 General If not otherwise specified, as a minimum analytical-grade chemicals shall be used; water shall be distilled or of a corresponding purity. “Solution“ shall be under
27、stood as an aqueous solution unless otherwise specified. 4.2 Methanol, CAS number: 67-56-1. 4.3 Dichloromethane, CAS number: 75-09-02. 4.4 Acetonitrile, CAS number: 75-05-8. 4.5 Ethylenediaminetetraacetic acid (EDTA) disodium salt dihydrate (Na2EDTA 2H2O), CAS number: 6381-92-6. 4.6 Oxalic acid dihy
28、drate, CAS number: 6153-56-6. 4.7 Acetic acid (glacial), CAS number: 64-19-7, mass fraction w = 99,8 g/100 g. 4.8 Acetic acid, molar concentration c = 0,02 mol/l. Weigh 1,20 g of acetic acid glacial (4.7) into a 1-l-volumetric flask and fill with water up to the calibration mark. 4.9 Methanol/acetic
29、 acid mixture Mix 80 parts by volume of methanol (4.2) and 20 parts by volume of acetic acid (4.8). BS EN 16342:2013EN 16342:2013 (E) 6 4.10 Sodium hydroxide solution, molar concentration c = 1 mol/l. 4.11 Eluents 4.11.1 Eluent A: 0,002 7 mol/l of oxalic acid dihydrate (4.6) and 0,001 mol/l of EDTA
30、(4.5) in water, pH 4,0: Pre-dissolve 0,37 g of EDTA in water, add 0,35 g of oxalic acid dihydrate (0,025 % of oxalic acid) and adjust to pH = 4,0 with sodium hydroxide solution (4.10) using a pH-Meter. Then fill with water up to the 1000 ml mark. 4.11.2 Eluent B: Acetonitrile (4.4). 4.12 Reference s
31、ubstances 4.12.1 General For the reference substances, no purity is defined. However, the purity of the reference substances shall be known to determine the definite amount of standard in the calibration solution. 4.12.2 Zinc pyrithione, CAS number: 13463-41-7. 4.12.3 Piroctone olamine, (1-hydroxy-4
32、-methyl-6-(2,4,4-trimethylpentyl)-2-pyridone), CAS number: 68890-66-4. 4.12.4 Climbazole, (1-(4-chlorophenoxy)-1-(imidazol-1-yl)-3,3-dimethyl-2-butanone), CAS number: 38083-17-9. 4.13 Stock solutions 4.13.1 General Fresh stock solutions need to be prepared each working day. 4.13.2 Zinc pyrithione st
33、ock solution, mass concentration = 250 mg/l. Weigh approximately 25 mg of zinc pyrithione (4.12.2) to the nearest 0,1 mg into a 100-ml-volumetric flask, dissolve in 50 ml of dichloromethane (4.3) and fill up to the mark with the methanol/acetic acid mixture (4.9). 4.13.3 Piroctone olamine and climba
34、zole stock solution, mass concentration = 250 mg/l. Weigh approximately 25 mg of each piroctone olamine (4.12.3) and climbazole (4.12.4) to the nearest 0,1 mg into a 100-ml-volumetric flask and fill up to the mark with the methanol/acetic acid mixture (4.9). 4.14 Calibration solutions Fresh calibrat
35、ion solutions shall be prepared each working day. The following scheme in Table 1 for the preparation of the calibration solutions has proved useful in practice. Together, the given amounts of the stock solutions in ml, are pipetted into 25-ml-volumetric flask and filled up to the mark with the meth
36、anol/acetic acid mixture (4.9). For calculation the concentration of the calibration solutions have to be corrected with the known purity of the reference substances. BS EN 16342:2013EN 16342:2013 (E) 7 Table 1 Calibration solution Calibration solution Zinc pyrithione (4.13.2) ml Piroctone olamine +
37、 Climbazole (4.13.3) ml Concentration g/ml 1 0,5 0,5 5 2 1 1 10 3 2 2 20 4 3 3 30 5 5 5 50 5 Apparatus and equipment 5.1 Analytical balance, with a precision of 0,1 mg. 5.2 Membrane filter, for solvent filtration, 0,45 m pore size. 5.3 Ultrasonic bath, with temperature controlled heater. 5.4 Disposa
38、ble syringes 5.5 Membrane filter, for sample filtration, e.g. PTFE, 0,45 m pore size. 5.6 High-performance liquid chromatograph, consisting of: sampling device; pump system with gradient function; degasser (optionally; eluent may be degassed prior to use if the system requirements are fulfilled (q.v
39、.); column oven; photodiode array detector (for quantitative determination without identification a multiple wavelength detector is sufficient); evaluation system. 5.7 Analytical reversed-phase separation column, e.g.: Onyx Monolithic C18 100 mm 3 mm (Phenomenex)1)or Chromolith RP18e, 100 mm 3 mm (M
40、erck)1). A pre-column packed with stationary phase similar to the analytical separation column shall be used. 6 Sampling The sampling technique is not part of the technique specified in the standard method. 1) This is an example of a suitable product available commercially. This information is given
41、 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. BS EN 16342:2013EN 16342:2013 (E) 8 7 Procedure 7.1 Sample preparation Weigh approximately 250 mg of sample
42、to the nearest 0,1 mg into a 50-ml-volumetric flask. Add 5 ml of dichloromethane (4.3) and 5 ml of methanol (4.2). Place the volumetric flask into a temperature controlled ultrasonic bath for 10 min and heat gently at 35 C to 40 C. When the sample is dissolved or homogeneously dispersed, let the sam
43、ple cool down at room temperature. Fill the volumetric flask to the mark with methanol/acetic acid (4.9) and shake. Filter approximately 2 ml of methanol/acetic acid mixture through a membrane filter (5.5) into a HPLC vial, discarding the first 0,5 ml. Fresh sample solutions shall be prepared each w
44、orking day. 7.2 High-performance liquid chromatography (HPLC) When starting measurements, examine the baseline stability and response linearity of the detector. The detector shall be able to detect the lowest calibration solution of climbazole (5 g/ml) at a signal to noise ratio of 6:1. The same ope
45、rating conditions of the HPLC System shall be maintained throughout the measurements of all sample and calibration solutions. When using the apparatus (5.6) and the column of (5.7), the following conditions have shown useful: flow: 2,0 ml/min injection volume: 5 l injector temperature: room temperat
46、ure column temperature: 30 C detection: Zinc pyrithione: detection wavelength: = 340 nm Climbazole: detection wavelength: = 277 nm Piroctone olamine: detection wavelength: = 305 nm running time: 6 min 7.3 Gradient elution Eluent A: 0,002 7 mol/l of oxalic acid + 0,001 mol/l of EDTA in water, pH 4,0
47、(4.11.1). Eluent B: Acetonitrile (4.4). For the gradient elution eluents A and B are used in accordance with the volume ratios and time intervals given in Table 2, the following conditions have proved useful: BS EN 16342:2013EN 16342:2013 (E) 9 Table 2 Gradient programme Time min Fraction eluent A %
48、 Fraction eluent B % 0 85 15 2 50 50 3,5 50 50 3,6 20 80 4,6 20 80 4,7 85 15 6 85 15 It is recommended to do a blank run after each 10 sample-measurements to detect any carry-over which may have occurred. For this purpose make a solution according 7.1 without sample. 8 Evaluations 8.1 Identification
49、 The anti-dandruff agents to be determined are identified by comparing the retention times of the sample with the calibration solutions and by comparing the spectra of the sample and the reference. A background correction shall be made if required. 8.2 Quantitative determination and calculation Quantification is performed by means of linear regression based on the peak areas or peak heights of the external standards. The calibration curves shall be rectilinear and the correlation coefficient should be at least 0,996 or
