1、 g49g50g3g38g50g51g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58pharmaceutical use Part 4: Biological requirements and test methodsThe European Standard EN ISO 887
2、1-4:2006 has the status of a British StandardICS 11.040.20Elastomeric parts for parenterals and for devices for BRITISH STANDARDBS EN ISO 8871-4:2006BS EN ISO 8871-4:2006This British Standard was published under the authority of the Standards Policy and Strategy Committee on 29 September 2006 BSI 20
3、06ISBN 0 580 49229 XAmendments issued since publicationAmd. No. Date Commentscontract. Users are responsible for its correct application.Compliance with a British Standard cannot confer immunity from legal obligations. National forewordThis British Standard was published by BSI. It is the UK impleme
4、ntation of EN ISO 8871-4:2006.The UK participation in its preparation was entrusted to Technical Committee CH/212, IVDs.A list of organizations represented on CH/212 can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a EUROPEAN ST
5、ANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 8871-4June 2006ICS 11.040.20 Supersedes EN ISO 8871:1997 English VersionElastomeric parts for parenterals and for devices forpharmaceutical use - Part 4: Biological requirements and testmethods (ISO 8871-4:2006)lments en lastomre pour administration parentr
6、ale etdispositifs usage pharmaceutique - Partie 4: Exigencesbiologiques et mthodes dessais (ISO 8871-4:2006)Elastomere Teile fr Parenteralia und fr Gerte zurpharmazeutischen Verwendung - Teil 4: BiologischeAnforderungen und Prfverfahren (ISO 8871-4:2006)This European Standard was approved by CEN on
7、5 June 2006.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obt
8、ained 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 translationunder the responsibility of a CEN member into its own language and notified to the Central Secre
9、tariat has the same status as the officialversions.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, Portug
10、al, Romania,Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2006 CEN All rights of exploitation in any form and by any means rese
11、rvedworldwide for CEN national Members.Ref. No. EN ISO 8871-4:2006: EForeword This document supersedes EN ISO 8871:1997. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the followingcountries are bound to implement this European Standard: Austria, Belgium,
12、Cyprus, Czech Republic, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. Endorsement notice This document (EN ISO 8871-4:2006) has been prepared by Technical Committee ISO/TC 76 “Transfusion, infusion and injection
13、equipment for medical and pharmaceutical use“ in collaboration with CMC. 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 December 2006, and conflicting national standards shall be withdrawn at the
14、 latest by December 2006. Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, The text of ISO 8871-4:2006 has been approved by CEN as EN ISO 8871-4:2006 without any modifications. EN ISO 8871-4:2006Reference numberISO 8871-4:2006(E)INTERNATIONAL S
15、TANDARD ISO8871-4First edition2006-06-15Elastomeric parts for parenterals and for devices for pharmaceutical use Part 4: Biological requirements and test methodslments en lastomre pour administration parentrale et dispositifs usage pharmaceutique Partie 4: Exigences biologiques et mthodes dessai EN
16、ISO 8871-4:2006ii iiiForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a
17、subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission
18、 (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical com
19、mittees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not
20、 be held responsible for identifying any or all such patent rights. ISO 8871-4 was prepared by Technical Committee ISO/TC 76, Transfusion, infusion and injection equipment for medical and pharmaceutical use. This first edition, together with parts 1, 2, 3 and 5, cancels and replaces ISO 8871:1990 an
21、d ISO 8871:1990/Amd.1:1995, which has been technically revised. ISO 8871 consists of the following parts, under the general title Elastomeric parts for parenterals and for devices for pharmaceutical use: Part 1: Extractables in aqueous autoclavates Part 2: Identification and characterization Part 3:
22、 Determination of released-particle count Part 4: Biological requirements and test methods Part 5: Functional requirements and testing EN ISO 8871-4:2006iv Introduction The pharmaceutical industry requires, to an increasing extent, concrete details from the rubber manufacturer about the biological s
23、tatus of rubber closures as far as elastomeric closures are used as primary packaging materials in direct contact with the medicinal products. This request has been taken into account by preparing Annexes A to D of this part of ISO 8871. Tests presented in this part of ISO 8871 can be taken into acc
24、ount as a guideline if the question of biological safety arises in context with primary packaging materials for pharmaceutical products. The use of certain tests of Annex A to Annex D in case of special applications of the packaging material should be agreed upon between users and manufacturers. EN
25、ISO 8871-4:20061Elastomeric parts for parenterals and for devices for pharmaceutical use Part 4: Biological requirements and test methods 1 Scope This part of ISO 8871 specifies biological requirements for elastomeric parts for parenterals and for devices for pharmaceutical use. It also specifies th
26、e test methods, i.e. it offers the extraction procedures for elastomeric parts, and it makes reference to relevant biological test instructions in Pharmacopoeias and standards. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated
27、 references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 10993-5, Biological evaluation of medical devices Part 5: Tests for in vitro cytotoxicity USP, The United States Pharmacopeia, United States Phar
28、macopeial Convention, Inc., Rockville, MD, USA 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 bacterial endotoxins lipo-polysaccharides from gram-negative bacteria 3.2 bioburden population of viable microorganisms on or in product and/or a p
29、ackage ISO 11737-1:, definition 3.1 3.3 cytotoxicity biological response of mammalian cell cultures in vitro using appropriate biological parameters to extracts of elastomeric parts 3.4 intracutaneous toxicity local response to extracts of elastomeric parts after intracutaneous injections into rabbi
30、ts EN ISO 8871-4:20062 3.5 systemic toxicity systemic response to extracts of elastomeric parts after injection into mice 4 Biological requirements 4.1 General The elastomeric parts shall not release any substances that may adversely affect the therapeutic effectiveness and safety of the pharmaceuti
31、cal solution, including those substances which can exhibit toxic or pyrogenic reactions. Selection of tests and their interpretation to be used in biological evaluations should take into account the chemical composition of the materials, including the conditions of exposure as well as the nature, de
32、gree, frequency and duration of exposure of the material. Cytotoxicity, systemic toxicity and intracutaneous toxicity shall be considered for type testing as material characteristics. Endotoxins and bioburden shall be considered for monitoring purposes. 4.2 Extractable bacterial endotoxins The limit
33、, specified as endotoxin units per square centimetre (EU/cm2) of elastomeric part or endotoxin units per millilitre (EU/ml) extractable endotoxins, shall be agreed upon between supplier and user. The test method shall be validated. NOTE Annex A includes an example of a test method for the determinat
34、ion of extractable bacterial endotoxins that can be used. 4.3 Bioburden The limit, specified as colony-forming units per square centimetre (cfu/cm2)or cfu per elastomeric part, shall be agreed upon between supplier and user. The bioburden method shall be validated. NOTE ISO 11737-1 can be used as a
35、guideline to establish and validate the method. 4.4 Toxicity 4.4.1 General Materials shall be tested in vitro for cytotoxicity in accordance with 4.4.2. Materials that meet the requirements of this test are not required to undergo further testing. Materials that do not meet the requirements of this
36、test shall be tested in vivo for systemic and intracutaneous toxicity in accordance with 4.4.3 and 4.4.4 respectively. 4.4.2 Cytotoxicity The test shall be performed according to Annex B. For assessment refer to the requirements specified in USP, chapter , “Biological Reactivity Tests, In Vitro”. EN
37、 ISO 8871-4:200634.4.3 Intracutaneous toxicity The test shall be performed according to Annex C. For assessment, refer to the requirements specified in USP, chapter , “Biological Reactivity Tests, In Vivo, Intracutaneous Test”. 4.4.4 Systemic toxicity The test shall be performed according to Annex D
38、. For assessment, refer to the requirements specified in USP, chapter . EN ISO 8871-4:20064 Annex A (informative) Test for extractable bacterial endotoxins A.1 General This Annex defines a conventional method for the extraction of bacterial endotoxins from the surface of elastomeric parts. The extra
39、ctable bacterial endotoxins are determined according to the methods specified in USP, chapter , “Bacterial Endotoxins Test” or Ph.Eur.2, chapter 2.6.14 “Bacterial Endotoxins”. A.2 Principle The extraction is performed by shaking elastomeric parts in endotoxin-free water. The determination of extract
40、able bacterial endotoxins is performed according to the methods specified in USP or Ph.Eur. using limulus amebocyte lysate (LAL) which has been obtained from horseshoe crab, Limulus polyphemus, and which has been prepared and characterized for use as an LAL reagent. A.3 Reagents and materials A.3.1
41、Depyrogenated glassware/heat stable instruments (e.g. forceps). A.3.2 Endotoxin-free water. A.3.3 Endotoxin reagents as specified in USP or Ph.Eur. A.4 Preparation of depyrogenated glassware and heat stable instruments A.4.1 Clean the glassware in a laboratory dish washer. Ensure that residual soap
42、is completely removed. A.4.2 Wrap the glassware in aluminium foil as follows: completely wrap test tubes; wrap only the neck part of sample containers; completely wrap forceps. A.4.3 Depyrogenate the wrapped parts at appropriate conditions, e.g. at least 30 min at 250 C. A.4.4 After depyrogenation,
43、keep the parts in the aluminium foil. A.5 Extraction of the elastomeric parts Aseptically transfer a number of elastomeric parts corresponding to a surface of (100 10) cm2into a depyrogenated sample container. Carefully add 100 ml endotoxin-free water, protect with cover film and shake (orbital shak
44、ing) for 5 min at room temperature. EN ISO 8871-4:20065If the surface area of (100 10) cm2cannot be achieved, add (1 0,1) ml for every additional square centimetre. In case of elastomeric parts with barely accessible cavities, e.g. 13 mm freeze drying closures, flashballs, needle shields etc., it ma
45、y be necessary to cut the parts before extraction in order to obtain adequate recoveries. If parts are cut, take into account the original surface area of the parts. A.6 Expression of results Express the results as EU/cm2elastomeric part or EU/ml extractable endotoxins. EN ISO 8871-4:20066 Annex B (
46、normative) Test for cytotoxicity B.1 Principle The following test is designed to determine the biological reactivity of mammalian cell cultures following contact with extracts of elastomeric parts. For details of performing the biological test itself, see USP, chapter or ISO 10993-5. B.2 Preparation
47、 of extract Prepare as specified for preparation of extracts in USP, chapter , or in ISO 10993-5 using serum supplemented mammalian cell culture medium. Extract with a sample/medium ratio of 25 cm2per 20 ml for 24 h at (37 1) C. B.3 Procedure Follow the instructions specified in USP, chapter or in I
48、SO 10993-5. B.4 Expression of results B.4.1 Express the results as follows: cytotoxic; non-cytotoxic. B.4.2 The sample is considered to be non-cytotoxic if the following criteria have been met: u 50 % of the cells are round and devoid of intracytoplasmic granules; no extensive cell lysis and empty a
49、reas between cells. EN ISO 8871-4:20067Annex C (normative) Test for intracutaneous toxicity C.1 Principle The following test is designed to determine local responses and skin irritations to extracts of elastomeric parts by intracutaneous injections into rabbits. For details of performing the biological test itself, see USP, chapter . C.2 Preparation of extract Prepare as specified for preparation of extracts in USP, chapter using 9 g/l NaCl solution. C.3 Procedure Follow the
