1、December 2014Translation by DIN-Sprachendienst.English price group 9No 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、 11.200!%Zj“2275571www.din.deDDIN ISO 29941Condoms Determination of nitrosamines migrating from natural rubber latexcondoms (ISO 29941:2010),English translation of DIN ISO 29941:2014-12Kondome Bestimmung von aus Naturkautschuklatexkondomen migrierenden Nitrosaminen(ISO 29941:2010),Englische bersetzu
3、ng von DIN ISO 29941:2014-12Prservatifs Dosage des nitrosamines migrant des prservatifs en latex de caoutchouc naturel(ISO 29941:2010),Traduction anglaise de DIN ISO 29941:2014-12www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 14 pages1
4、2.14 2A comma is used as the decimal marker. Contents Page National foreword .3 National Annex NA (informative) Bibliography .4 1 Scope .5 2 Normative references .5 3 Terms and definition.5 4 Principle .5 5 Reagents and materials .6 6 Apparatus 7 7 Procedure 9 8 Calculation and evaluation of the res
5、ults of the N-nitrosamine content of the solution 11 9 Confirmation of the detected N-nitrosamines 12 10 Test report 13 Bibliography . 14 DIN ISO 29941:2014-12National foreword The text of this document (ISO 29941:2010) has been prepared by Technical Committee ISO/TC 157 “Non-systemic contraceptives
6、 and STI barrier prophylactics”. The responsible German body involved in its preparation was the DIN-Normenausschuss Medizin (DIN Standards Committee Medicine), Working Committee NA 063-01-04 AA Kondome und Intrauterinpessare. Attention is drawn to the possibility that some of the elements of this d
7、ocument may be the subject of patent rights. DIN and/or DKE shall not be held responsible for identifying any or all such patent rights. In addition to the legal units of measurement, this standard also uses the unit “inch”. It should be noted that the Gesetz ber Einheiten im Messwesen (German Law o
8、n units in metrology) prohibits the use of this unit for official and commercial purposes in Germany. The indication of this unit solely serves to facilitate the communication with those countries where this unit is used (import-export business). Conversion: Non-SI unit SI unit Conversion Subscript
9、in (inch) mm 1 inch = 25,4 mm “ The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 3696 DIN ISO 3696 ISO 4074 DIN EN ISO 4074 ISO/IEC 17025 DIN EN ISO/IEC 17025 3DIN ISO 29941:2014-12National Annex NA (informative) Bibliography DIN ISO 369
10、6, Water for analytical laboratory use Specification and test methods DIN EN ISO 4074, Natural latex condoms Requirements and test methods DIN EN ISO/IEC 17025, General requirements for the competence of testing and calibration laboratories 4DIN ISO 29941:2014-12Condoms Determination of nitrosamines
11、 migrating from natural rubber latex condoms 1 Scope This International Standard specifies a test method to determine the release of N-nitrosamines from condoms made from natural rubber latex. The method can also be used for other products such as probe covers, prophylactic dams, female condoms and
12、condoms made from synthetic materials, although there was no experience of testing such products at the time of publication of this International Standard. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the
13、 edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 3696, Water for analytical laboratory use Specification and test methods ISO 4074, Natural rubber latex condoms Requirements and test methods ISO/IEC 17025, General r
14、equirements for the competence of testing and calibration laboratories 3 Terms and definition For the purposes of this document, the following terms and definitions apply. 3.1 condom medical device used by consumers, which is intended to cover and be retained on the penis during sexual activity, for
15、 purposes of contraception and prevention of sexually transmitted infections NOTE If a consumer could responsibly consider a device to be a condom (due to its shape, packaging, etc.), it is considered a condom for the purpose of this International Standard. 4 Principle WARNING Owing to their toxicit
16、y, N-nitrosamines can be detrimental to human health. The testing laboratories should take special care to observe the occupational health and safety standards. N-nitrosamines are extracted into water. After having been concentrated, the water is tested for its N-nitrosamine content by means of gas
17、chromatography (GC) using a chemiluminescence detector. The test is performed in an environment that is free of volatile N-nitrosamines. The released N-nitrosamines are given in nanograms per grams (ng/g) of the sample. 5DIN ISO 29941:2014-125 Reagents and materials IMPORTANT Owing to the fact that
18、N-nitrosamines are decomposed by UV light, the exposure to sunlight or fluorescent light of the standardized solutions during the preparation should be avoided. Standardized solutions, migrate solutions and water should be protected by means of aluminium foil and stored in the dark at temperatures b
19、elow 5 C. Use only reagents of recognized analytical grade and only water conforming to ISO 3696, grade 3, unless otherwise specified. 5.1 Dichloromethane, which should be checked for the absence of N-nitrosamines. 5.2 Diatomaceous earth, from liquid-liquid extraction, with a specific surface area o
20、f 1 m2/g, a pore size of 3 000 nm to 8 500 nm and a particle size of 150 m to 650 m. Heat for 1 h at 200 C and wash with dichloromethane (5.1) after cooling. Another separating agent can be used providing it has been validated against diatomaceous earth. 5.3 n-Hexane. 5.4 Sodium hydroxide solution i
21、n water, c(NaOH) = 1 mol/l. 5.5 Nitrogen, with a volume fraction of at least 99,996 %. 5.6 Boiling chips. 5.7 Sintered glass frit for columns (6.3 and 6.4). 5.8 Acetone. 5.9 N-nitrosamine standardized solutions. Prepare standardized solutions with known quantities of the N-nitrosamines to be determi
22、ned in n-hexane (5.3) in the concentration range of 100 ng/l to 300 ng/l. The following N-nitrosamines are of importance for condoms from elastomer or rubber. The list is, however, not comprehensive: N-nitrosodimethylamine (NDMA); N-nitrosodiethylamine (NDEA); N-nitrosodibutylamine (NDBA). If there
23、are indications of the presence of other N-nitrosamines relevant for toxicology or if the nature of the used vulcanization accelerators supports this indication these other N-nitrosamines should also be tested, examples could be: N-nitrosodibenzylamine (NDBzA); N-nitrosodiisononylamine (NDiNA), i.e.
24、 N-nitroso-3,3,5-trimethylhexylamine. 5.10 Solution of the internal standard of 200 ng/ml N-nitrosodiisopropylamin (NDiPA) in acetone (5.8). The solution shall be free of other N-nitrosamines. 6DIN ISO 29941:2014-125.11 Water-free sodium sulfate, granulated, or a suitable phase separation filter for
25、 the Whatman apparatus. Pre-wash 30 g of sodium sulfate with 25 ml of dichloromethane (5.1). 5.12 Ammonia solution in water, c(NH3) = 0,1 mol/l. 5.13 Sea sand, washed with acid and calcined. 6 Apparatus Usual laboratory apparatus and, in particular, the following: 6.1 Glass vessels washed with acidi
26、c cleansing agents, shall be treated with ammonia solution (5.12), rinsed with water and dried before being used. 6.2 Drying cabinet, adjustable to (40 2) C. 6.3 Glass column, with outlet and plug made from polytetrafluorethylene (PTFE). Length of the column: (300 10) mm, inner diameter: (26 1) mm.
27、6.4 Glass column, with outlet and plug made from polytetrafluorethylene (PTFE). Length of the column: (300 10) mm, inner diameter: (15 1) mm. 6.5 Kuderna-Danish vaporizer, modified with a graduated collecting vessel and an air cooler. An alternative apparatus may be used provided that its performanc
28、e has been validated against the Kuderna-Danish vaporizer. 6.6 Water bath, temperatures adjustable from 40 C to 60 C. 6.7 Vials, closable with vial mouth rings and N-nitrosamine free or PTFE coated septa. 6.8 Pliers, for closing the vials (6.7). 6.9 Fibreglass plug, washed with dichloromethane (5.1)
29、. 6.10 200 ml separating funnel. 6.11 100 ml separating funnel. 6.12 Chemiluminescence detector, of sufficient sensitivity Thermal Energy Analyzer, TEA1). Another analysis detector may be used providing it has been validated against the TEA. 6.13 Chromatography system. The decision regarding the sel
30、ection of the chromatography system may be made by the tester. The test laboratory shall, however, provide evidence that the conditions were optimized in such a way as to achieve a sufficient separation of peaks, with the following points being observed: The system shall separate the N-nitrosamines
31、(5.9) mentioned in this International Standard such that their peak areas can be compared with the peak area of the internal standard solution (5.10). The system separates N-nitrodimethylamine and N-nitrodiethylamine from the mentioned N-nitrosamines. 1) Thermal Energy Analyzer is an example of a su
32、itable product available commercially. This information is given for the convenience of the users of this International Standard and does not constitute an endorsement by ISO of this product. 7DIN ISO 29941:2014-12It is possible that two separating columns are necessary to separate all the N-nitrosa
33、mines and to achieve a sufficient sensitivity for NDBzA, if it is part of the test. The following conditions have been found to be suitable for the determination of volatile N-nitrosamines when using gas chromatography. EXAMPLE 1 Packed columns Injector temperature: 200 C Oven temperature: 200 C Col
34、umns: 2,5 m to 3,0 m glass, outer diameter 1/8“, packed with: 15 % Carbowax 20 M, TPA on Chromosorb WHP 100/120 mesh2); or 10 % Carbowax 20 M, 2 % KOH on Chromosorb WHP 100/120 mesh; or 4,0 m to 5,0 m glass, outer diameter 1/8“, packed with: 15 % SP 1220, 1 % H3PO4on Chromosorb WAW 100/120 mesh. Tem
35、perature of the pyrolysis oven: 480 C Carrier gas: Argon, helium, or nitrogen with a flow rate of (20 1) ml/min Coupling: Directly between gas chromatography column and pyrolysis oven or by applying a coupling heated to 250 C. As regards the determination of alkylphenyl-N-nitrosamines, the following
36、 conditions have been found to be suitable: Injector temperature: 150 C. Oven temperature: between 120 C and 130 C. Columns: 2 m glass, outer diameter 1/4“, inner diameter 2,0 mm, packed with: 10 % OV-101 on Chromosorb WHP 80/100 mesh; or 3 % OV-225 on Chromosorb WHP 80/100 mesh. Temperature of the
37、pyrolysis oven: 480 C Temperature of the coupling: 250 C. EXAMPLE 2 Capillary columns Either Injector temperature: 200 C Oven temperature: 60 C, 230 C (10 C/min) Column: 25,0 m quartz capillary 0,53 mm FFAP 1 m 2) Carbowax and Chromosorb are examples of suitable products available commercially. This
38、 information is given for the convenience of the users of this International Standard and does not constitute an endorsement by ISO of these products. 8DIN ISO 29941:2014-12Temperature of the pyrolysis oven: 480 C Temperature of the coupling: 250 C or Injector temperature: 50 C, 1 min 200 C (75 C/mi
39、n) Oven temperature 40 C, 7 min 60 C (1 C/min), 230 C (14 C/min) Column: 30,0 m quartz capillary, 0,53 mm, SE-54-film of 2 m Temperature of the pyrolysis oven: 480 C Temperature of the coupling: 250 C. 7 Procedure 7.1 Migration of the condom material 7.1.1 For this test, use only condoms that have b
40、een tested according to ISO 4074 and conform to that standard. Use the average results of the tests for length and width for the calculations (see Clause 8). Determine the masses of three condoms. The average mass of these three reference condoms is the basis for the quantity of intact condoms to be
41、 used in the test. Cut each condom in half. Weigh (5 1) g of the prepared condoms to the nearest 0,05 g and transfer them to a 50 ml Erlenmeyer flask. Add 40,0 ml of water by means of a pipette. Close the flask by means of a glass stopper, shake it cautiously so that the solution covers the prepared
42、 condoms. Store the flask in the drying cabinet (6.2) for 10 min ( 30 s) at (40 2) C. Adapt the reagent quantities and the apparatus sizes proportionally if the mass of the condom material exceeds 5 g. The quantity of added internal standard solution (5.10), however, is always 1,0 ml. 7.1.2 Decant t
43、he solution from the flask to a 50 ml measuring cylinder with glass stopper, wash the condom material with 4,0 ml of water, add it to the test solution, fill with water up to the mark and mix it for at least 1 min. 7.2 Isolation of the N-nitrosamines in the solution 7.2.1 General Add 1,0 ml of the i
44、nternal standard solution (5.10) and 1,0 ml of sodium hydroxide solution (5.4) to the solution in the measuring cylinder (7.1.2) by means of a pipette. The test solution can be processed according to procedure A or B. 7.2.2 Procedure A 7.2.2.1 Fill, with (25,0 0,1) g of diatomaceous earth or another
45、 suitable separating agent (5.2), the glass column inner diameter of 26 mm; the bottom of which is closed by means of a fiberglass plug (6.9). Cover the top of the column with the sintered glass frit (5.7) or with a sand layer of 1 cm (5.13). When filling the column, tap the outside gently to ensure
46、 an even distribution. 9DIN ISO 29941:2014-127.2.2.2 Close the measuring cylinder with the solution (7.2.1) and shake it until it is homogeneous. Then, slowly transfer it to the prepared column with diatomaceous earth or equivalent material (7.2.2.1). The test solution shall be distributed as statio
47、nary phase onto the porous matrix within a period of 10 min to 15 min. In the lower part of the column, there remains a dry zone of a width of between 50 mm and 70 mm. 7.2.2.3 Pour 60 ml to 80 ml of dichloromethane onto the column, collect 40 ml of eluate in the Kuderna-Danish collecting vessel or a
48、n equivalent vessel (6.5) within a period of 15 min to 25 min, with the dropping speed being regulated by means of the PTFE plug. NOTE During the elution with dichloromethane, the dry zone will be decreased to 15 mm to 30 mm. This process can be well observed because of the different tinges of the d
49、iatomaceous earth zone that is wet from the test solution and the diatomaceous earth zone that is wet from dichloromethane. It is important not to reach the capacity limit of this dry zone; otherwise, the test solution might contain water. 7.2.3 Procedure B 7.2.3.1 Close the measuring cylinder with the solution (7.2.1), shake it until it is homogeneous; slowly pour the solution into the separating funnel (6.10
