1、BRITISH STANDARD Biotechnology - Equipment - Guidance on testing procedures for cleanability BS EN 12296:1998 The European Standard EN 122961998 has the status of a British Standard ICs 07.080; 07.100.01 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW CAa-Z STD-BSI BS EN 12296
2、-ENGL 1998 m Lb24669 0716156 640 m direction of the Sector Board for Amd. No. Date Materiais and Chemicals, was published under the authority of the Standards Board and comes into effect on 15 August 1998 BS EN 12296:1998 Text affected National foreword This British Standard is the English language
3、version of EN 122961998. The UK participation in its preparation was entnisted to Technical Committee CII/58, Biotechnology, which has the responsibility to: - aid enquirers to understand the text; - present to the responsible international European committee any enquiries on the interpretation, or
4、proposals for change, and keep the UK interests informed; them in the UK. - monitor related international and European developments and promulgate A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement
5、 international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitied “International Standards Correspondence Index“, or by using the “Find“ facility of the BSI Standards Electxonic Catalogue. A British Standard does not purport t
6、o include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover,
7、 the EN title page, pages 2 to 7 and a back cover. O BSI 1998 I l ISBN O 680 30067 6 STD.BS1 BS EN 12296-ENGL 1998 m 1b24bb 071b157 587 m EN 12296 March 1998 ICs Descriptors: Biotechnoiom, medical equipment, cleaning, disinfection, contamination, micro-organisms, noxious micro-organisms, tests, safe
8、ty, hygiene conditions, inspection, accident prevention, environmental protection, work safety Engush version BiotechnooloB - Equipment - Guidanee on testing procedures for cleanability Biotechnologie - Equipement - Guide des procdures dessai pour le contrle de la capacit au nettoyage Reinigbarkeit
9、Biotechnik - Gerate und Ausrustun gen - Leitfaden fr Verfahren zur Prufung der This European Standard was approved by CEN on 2 March 1998. CEN members are bound to comply with the CENKENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a nationa
10、l standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained 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 o
11、ther language made by translation under the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany,
12、 Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. CEN European Committee for Standardization Comit Europen de Normalisation Europisches Komitee fr Normung Central Secretariat: rue de Stassart 36, B-1050 Brussels O 1998 CEN All
13、 rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 122961998 E STD=BSI BS EN 1229b-ENGL L998 m 1624bb9 07LbL58 413 m Page 2 EN 12296:1998 For e w or d This European Standard has been prepared by Technical Committee CENliC 233, Biotechnology,
14、 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 text or by endorsement, at the latest by September 1998, and conflicting national standards shd be withdram at the latest by September 1998. Acco
15、rdmg to the CEN/CENELEC Internal ReguLations, the national standards organizations of the following countries are bound to implement this European Standard Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Porhigal,
16、 Spain, Sweden, Switzerland and the United Kingdom. Contents Foreword Introduction 1 Scope 2 Definitions 3 Testing 4 Documentation Annex A (informative) Selection guide on test methods for cleanllliess Annex B (informative) Information on test methods for cleanliness Annex C (informative) Bibliograp
17、hy we 2 3 3 3 4 4 5 6 7 O BSI 1998 Introduction The cleaning of plant and equipment is an essential element of biotechnology processes in order to protect the safety of people and the environment and to avoid harmful operational effects through the accumulation of soil. Testing procedures should be
18、developed and documented to ensure that relevant information on cleanability is available. Standards (e.g. EN IS0 9000 series, see annex C 9), guidelines (e.g. Good Manufacturing Practice (GMP) see annex C lo) state general procedures of good practice which facilitate high quality manufacturing if f
19、ollowed. This European Standard refers to assessing the cleanability of equipment used in biokchnoloa, where additional specific requirements related to safety and to special features of biotechnological processes are required. It should be read in association with the more geneml standards and guid
20、elines as mentioned above. in particular this European Standard states the principles on which test methodology is based. Informative guidance on selection of test methods is provided in annex k The extent to which it is necessary to remove soil from equipment and plant varies substantially with the
21、 process. In some cases abundant residues after cleaning do not ham people or the environment or do not cause difficulties in the process. In others very low residues are essential. The complete removal of soil on surfaces cannot be achieved, because for example all surfaces are adsorptive to some d
22、egree. 1 scope This European Standard gives guidance on general testing procedures to assess the cleanability of equipment (components and units of equipment) used in biotechnological processes. This European Standard applies primarily if the intended use of equipment includes the use of potentially
23、 hazardous micro-organisms. This European Standard also applies to non-hazardous micro-organisms andor to residual soil which can adversely affect sterilization processes or which can cause crosscontamination of products or processes. This European Standard applies to plants or components such as, v
24、alves and fitting, tanks, pumps, piping, separating and filling devices as well as instrumentation in contact with process fluids. Page 3 EN 12296.1998 2 Definitions For the purposes of this standard, the following definitions apply: 2.1 clean condition of (a) product, surface, device, gases and/or
25、liquids with residual soil below a defined threshold level 2.2 cleanability ability to be made clean 2.3 cleaning removal of soil 2.4 Cleaning-In Place (CIP) cleaning without dismantling of components of equipment andor unit of equipment 2.6 components of equipment technical entity which forms part
26、of a unit of equipment NOTE and sensors. 2.6 contamination presence of soil 2.7 residual soil soil left after cleaning 2.8 soil material, including micro-organisms, metabolites and components of process media present on a surface 2.9 unit of equipment assembly of components used to perform one or mo
27、re unit operations Examples of components of equipment are vessels, valves O ES1 1998 Page 4 EN 12296:1998 3 Testing 3.1 General The requirements for cleanability vary with process, especdy in relation to its assessed risk. Therefore cleanability classes for biosafety are defined in individual equip
28、ment standards. This will allow the manufacturers of plant and equipment to state the performance of their equipment using test methods, includuig visual inspection, developed according to the principles described in 3.2. It will also allow users of equipment to define their requirements in simple t
29、erms. These classes define performance only in relation to a defined indicator substance(s) and one or more defined cleaning protocol(s) and are relevant to the proposed use in the equipment. The cleanability class assigned to the equipment is likely to vary with the indicator and cleaning protocol
30、chosen. The soil adhering to surfaces at the end of a biotechnology process will contain many constituents. The indicator substance chosen to demonstrate the cleanability should be representative of those constituents that have an impact on safety aspects in relation to the need to protect people, t
31、he environment or features of the process. A brief description of indicators and test methods is given in annex B. 3.2 Methodology Tb determine the cleanability of plant and equipment, choose and specify an appropriate test method or combination of test methods (see annexes A and B): a) specify an a
32、ppropriate indicator related to the proposed use of the equipment; b) select the sampling procedure and the analytical procedure to be used to determine the quantity of this indicator which is present on relevant equipment surfaces or on surfaces in the plant ; c) specify a cleaning protocol includi
33、ng, as a minimum, the specification of the constituents of the cleaning material and the mode of application. NOTE 1 Potential hazards to the operator during cleaning should be assessed. NOTE 2 Factors such as the duration, temperature and fluid flow rates of cleaning should be included in the proto
34、col. NOTE 3 The cleaning protocol can consist of a number of successive operations. 3.3 Testing procedure Camy out the testing procedures as follows: a) load the equipment or plant with the indicator under normal operating conditions or in a way which simulates these; b) run the equipment andor plan
35、t under normal or simulated-normal conditions until the load containing the indicator has been discharged from the equipment and the equipment and/or plant is ready to be cleaned c) using the analytical procedure selected in 3.2, detennine the quantity of indicator substance present after discharge
36、of the load but before cleaning; d) apply the cleaning protocol specified in 3.2 to the plant or equipment being tested for clean f) using the data obtained, express the cleanability of the equipment or plant; g) determine the appropriate cleanability class to the equipment under test as described i
37、n the equipment standards with respect to the chosen indicator substance and cleaning protocol. NOTE The procedure described by European Hygienic Design criteria Group (EHEDG) (see annex C 2) can be quoted as an example of the application of 3.2 above, in which the indicator substance is the spores
38、of a specific bacterium, the analytical procedure is a culturing method which detects the number of this bacterium present before and after cleaning and in which the cleaning protocol is described (see annex C 3). Clearly other approaches can be used for the indicator substances could be another mic
39、roorganism, a specific member of a chemical group such as a defined protein, carbohydrate or lipid, a specinc compound known to be harmful to people or to the environment, or to future processing (see annex B). Potential cleaning protocols can be simple, for example a wash with water applied through
40、 a hosepipe or complex, as with sophisticated in-place cleaning involving the use of hazardous chemid at high temperature. Many test methods are possible, ranging from the use of a biological indicator to the use of chemical assays, immunological techniques, fluorescence assays and physical test met
41、hods, including microscopy (see annex B). 4 Documentation The equipment manufacturer/supplier andor the user should establish and document the procedure(s) used for the assessment of the cleanability of the component or unit of equipment. This documentation should include the applied test conditions
42、 (testhg method, indicator, analytical procedure and cleaning protocol) and the results of the test. O BSI 1998 STDnBSI ES EN 12296-ENGL L77B m 1624669 071blbl TOB Page 5 EN 122W.1998 Annex A (informative) Selection guide on test methods for cleanliness Fgure Al gives guidance on the selection of te
43、st methods for cleanability It represents a decision tree for selection of a cleanability test method based on scale, and access of the equipment. small large I access ? 6 1 2 3 4 * rinse (see B.4), visual inspection (see B.2), swab (see B.3), optional rinse, test following batch for contamination (
44、see B.5) rinse *, visual inspection, swab * rinse, test following batch for contamination optional compare results, if they are consistent sample rinse-fluid only Figure A.l- Decision tree for selection of a cleanability test method O BSI 1998 Page 6 EN 12296.1998 Annex B (informative) Information o
45、n test methods for cleanliness B.l Generai Essentially four test methods are available to determine the level of cleanliness of equipment (see annex C 3). They are visual inspection, swabbing the inner surfaces, sampling the final rinse and testing the following batch for contamination. B.2 Visual i
46、nspection Residual soil, adsorbed microbial populations or relevant mer indicators may be detected by a visual inspection of the equipment under investigation. This detection can be done by the eye or by using microscopic techniques such as light microscopy, scanning or transmission electron microsc
47、opy (see annex C 4). In most cases it wili be necessary to gain access to the equipment under investigation in order to be able to use these techniques. B.3 Swabbing the inner surfaces Indicator substances (chemical or (micro-)biological) can be measured by swabbing exactly defined areas of equipmen
48、t that come into contact with the product. Swabbing of easily accessible flat surfaces may cause overapthistic results with regard to the cleaning. Swabbing poorly accessible spots can produce over-pessimistic results. Vitally important for statistically relevant results are a correct choice of plac
49、es where to swab in the equipment and the area to be swabbed when using this test method Swabbing can be done either direct e.g. with agar count plates or indirect with e.g. cotton wool, wate tips or petsifilmS. ThLs test method also requires that access may be gained to the equipment under investigation. As an example, a very sensitive and simple and inexpensive method for the detection of residual amino acids and proteins on surfaces is described as follows. The ninhydrin method is based on the reaction of mino acids, peptides and proteins with tnketohydrindenhyd. Th
