1、STD-BSI BS EN 12b89-ENGL 1798 Lb24bb9 0737b03 499 BRITISH STANDARD Biotechnology - Guidance on assessment of the purity, biological activity and stability of micro-organism based products The European Standard EN 12689:1998 has the status of a British Standard ICs 07.080; 07.100.01 BS EN 12689: 1998
2、 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW - STD*BSI BS EN 12b89-ENGL 1998 1b24bb9 0737b04 325 H BS EN 126891998 D* National foreword Text affectai This British Standard is the English language version of EN 126891998. The UK participation in its preparation was entruste
3、d to Tkchnical Committee CW58, Biotechnology, which has the responsibility to: - aid enquirers to understand the text; - present to the responsible European committee any enquiries on the - monitor related international and European developments and promulgate interpretation, or proposals for change
4、, and keep the UK interests informed; them in the UK. A list of organizations represented on this cormnittee can be obtained on request to its secretary. Cross-references The British Standards which implement internalional or European publications referred to in this document may be found in the BSI
5、 Standarck Catalogue under the section entitled “international Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contra b) a specified level of biocontaminants;
6、c) identity of the microbial component to strains. 2.9 product stability preservation of biological activity andor purity over time and under defined conditions 2.10 reference data documentation of the characterization of micro-organism includmg any methods which are state-of-art O BSI 1998 STD-BSI
7、BS EN 12b7-ENGL 1778 lb24bb7 0737b08 T70 Page 4 EN 12689:1998 3 General considerations 3.1 Assessment protocol The assessment of technical specifications should be consistent with the intended use of the product. This assessment refers only to the microbial component or the biological activity of th
8、e products. Identification of the microbial component should be provided by the manufacturer to the user. “he mufacturer can use a wide range of procedures to characterize the microbial component, depending on the chamctecs of the micro-organism(s). The manufacturer should process the microbial comp
9、onent in such a way that its identity is ensured in the end product. NOTE Attention is drawn to EN 1619 regarding the strain conservation of micro-organisms. Validity of assessment of technical specifications is assured when specific criteria and factors are considered. The technical specifications
10、should be assessed using appropriate methods, depending on: - the micro-organism contained in the MBP - the purpose and design of the assessment. The method of assessment should be provided by the manufacturer. These methods should be used as the basis of the evaluation of product qual b) execution
11、of the assessment protocol (see 3.3); c) record keeping (see 3.4). 3.2 Design and review of the assessment protocol The design of the assessment protocol should be documented and reviewed for validity according the criteria in clauses 4,6 and 6. The assessment protocol should include procedures for
12、sampling and pretreatment of samples. The assessment of technical specifications is carried out on product samples as prepared for use. The sampling should be consistent with the type of applied statistical analysis. The type of applied stastical anaysis should provide representative data concerning
13、 production. NOTE Sampling methods and sample pretreatment methods described by national and international standards for the type of product to be tested should be used. Degradation and contamination of the MBP microbial component should be minimiied during storage and transportation of samples. Con
14、ditions of storage and transportation of MBP samples, such as temperature, humidity and light, should be as close as possible to those of the storage of the whole product. Pretreatment of samples is usually necessary for the isolation of the microbial component of the MBl? Pretreatment of samples is
15、 carried out after collection and storage if required. The choice of a pretreatment procedure depends on: - the nature and composition of the MBPs (e.g. liquid or solid samples); - the applied method to assess product puri, biological activity of the product and product stabiity. pre-tseatment can a
16、lter the number and activity of the microbial component of the MBPs. Therefore, it is essential that the pretreatment procedure should be validated before being incorporated into assessment protocols. 3.3 Execution of assessment protocol The assessment protocol should be carried out in accordance wi
17、th the design as follows: - COU - store the samples under conditions which minimize degradation of the anal; - bre-treat the samples; - record any deviation bm the assessment protQc01. - Collect data and Obtain results; 3.4 Record keeping A record should be kept of aii aspects of the assessment prot
18、ocol. This should not be considered an exhaustive list and should include: - identification of the test as foreseen by the assessment protocoi; - person performing the test; - date of the test; - method chosen for technical specification assessment; - origin, storage and transportation of samples; -
19、 number and size (weight andor volume) of samples tested; - controls used; - deviation from the protocol; - interpretation of the results. 4 Validity of purity assessment 4.1 General Purity assessment provides preliminary data on biological activity and dosage of the product. A iarge variety of MBPs
20、 exists and many methods are available to assess punty. For any type of product the validity of purity assessment depends on the criteria and factom given in 4.2 to 4.6. O BSI 1998 STD-BSI BS EN 12b89-ENGL 1998 = lb24bb9 0737b07 907 W 4.2 Identification of the microbial component Purity is expressed
21、 by quantitative data based on the evidence of the identity of the microbial component. Therefore, identity is the primary factor to be confirmed to validate purity assessment. NOTE Identification methods described by national and international standards for the type(s) of micro-organism to be ident
22、ined should be used. Identity can be expressed as the degree of morphological or genetic similarity of the micro-organism contained in the MBP to the reference strain(s). Identity can be determined from the analysis of factors related to the activity of the micro-organisms. Methods applied to identi
23、fy the microbial component such as serological, genetic and physiological analysis or isolation by use of selective media or other appropriate substrates should have been previously calibrated or validated. 4.3 Quantitative data Quantitative data on purity refers to a method of calculation and to th
24、e precision of results which should both be reported in the analysis of the results. 4.4 Availability of controls and/or reference data Availability of controls andor reference data improves the assessment of the applied method(s). MBP sample results should be compared with control results or with r
25、eference data provided by the manufacturer. Negative controls consist of a MBP sample without its microbial component. Negative Controls are processed under the same conditions as the MBP sample containing the microbial component. Negative controls are used to check for any contamination occurring d
26、uring execution of the assessment protocol. Positive controls can consist of a sample of the reference strain whose characteristics are known. When such a positive control is not available, the results of the assessment should be confirmed by comparison with reference data - the target for the micro
27、bial component such as living organisms, substances, plant tissue on which the MBP produces the desired effects; - the mechanism of interaction of the microbial component with its target. 6.2 Quantification of the biological activity of the product The quantification of the biological activity of th
28、e product by direct or comparative measurement refers to a method of calculation and to the precision of results which should both be reported in the anaiysis of results. 6.3 Availability of controls and/or reference data The avaiiabiity of controls andor reference data improves the assessment of th
29、e applied method(s). MBP sample results should be compared with control results or with reference data provided by the manufacturer. Negative controls consist of a MBP sample without its microbial component. Negative controls are processed under the same conditions as the MBP sample containing the m
30、icrobial component. Negative controls are used to check for any contamhation occurring during the execution of the assessment protocol. A positive control can consist of a sample of material that contains a comparable activiQ biological or chemical, of known characteristics. When such a positive con
31、trol is not available, the results of the assessment should be confirmed by comparison with reference data O BSI 1998 STD.BS1 BS EN L2bA7-ENGL 1778 Lb24bb7 0737bLO b27 Page 6 EN 12689:1998 6.4 Strategies for application of the MBPs Application conditions can affect microbial performance, especially
32、when the product is based on a mixture of micro-organisms. A careful consideration of product application conditions is recommended. 6.6 Biotic and abiotic factors Preliminary studies on environmental variables, which directly affect the biological activity of the product, improve the reliability of
33、 the designed assessment protocol. A careful consideration of such factors is recommended. 6.6 Mechanism involved in the product-environment interaction For the purpose of a product quality evaluation, the level of understanding for the mechanism involved in the product-environment interaction shoul
34、d be consistent with the intended use of the product. 6 Validity of stability assessment Product stability data indicate product purity andor the biological activity of the product as defined in clauses 4 and 6 over a period of time and under defined conditions. The aim of the stability assessment f
35、or product quality evaluation is to conirm, within acceptable limits, the MBP technid specifications over a period of time and under defined conditions such as the product storage period. Stability studies should take it into consideration that microbial degradahon or inactivation varies according t
36、o storage period and storage conditions. The following criteria and factors are hiportant to validate stability assessment. a) Stability should be expressed as the confirmation of change or lack of change of the biological activity of product and/or product purity at specific point of time. Environm
37、ental factors and storage conditions should be defined and controied Wherever significant changes or product degradation occur, acceptable limits should be defined with respect to the intended use of the product. b) The stability assessment should be carried out with respect to: - a period of time o
38、r the proposed shelf life; - established environmental factors or the proposed storage conditions. NOTE The temperature of the product can be the most cruciai factor, while factors such as humidity and light can be less important when it has been demonstrated that the container protects the product.
39、 c) When shelf life is proposed, the stability assessment should be conducted more frequently during the beginning of the shelf life period followed by longer testing intends towards the end of the period. d) Stability data of freezedried products can be obtained ar their reconstitution. O BSI 1998
40、Page 7 EN 12689:1998 Annex A (informative) Bibliography EN 1619, Biotechnology - Large s& process and production - Geneml requirements for management and organizatwn for strain conservation pcedures. IS0 4833, Microbiology - Geneml guidance for the enumeration of micro-organisms - Colony count techn
41、ique at 30 “C. IS0 6887, Microbiology - Geneml guidance for ULe preparatiun of dilutions for a microbiological examination. IS0 7218, Microbiology - Geneml guidance for microbiological examinations. Council Directive 90/219/EEC of 23 414 1990 on the contained use of genetically modified micro-organi
42、sms. OJEC 08.05.1990, No. L 117, p 1. Council Directive 90/220/EEC of 23 4ril 1990 on the deliberate release into the environment of genetically modified organisms. OJEC 08.05.1990, No. L 117, p 15. EN 12686, Biotechnology - Modvied organisms for application in the enwinmment - Guidance for the samp
43、ling strate for deliberate releases of genetically modifid micro-organisms, including wimes. EN 12682, Biotechnology - ModijYed organism for application in the enwimnment - Guidance for the characterization of genetically moded organh by analysis of the functional q&n of the genomic modification. EN
44、 12683, Biotechnologz/ - Modvied organisms for application in the environment - Guiunce for Uze characatitm of genetically modvied organism by analysis of u2R molecular stability of the genomic modification. 101 EN 12687, Biotechnology - Modified organisms for applicaticm in ULe environment - Guidan
45、ce for th? chamctemzation of genetically modified organism by aruysis of the gmic modification. li EN 12685, Biotechnology - Modvied organisms for application in the envinmment - Guidance for the monitoring strategies for deliberate releases of gmtixdy modified micro-organisms, including wiruses. i2
46、1 Council Directive 90/679/EEC of 26 November 1990 on the protection of workers from risks related to exposure to biological agents at work (seventh individual Directive within the meaning of Article 16 (1) of Directive 89/391/EEC). OJEC 31.12.1990, No. L374, p 1. O BSI 1998 S BS EN 12689:1998 BSI 3
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