Appendix XVI B. Microbiological Examination of Non-sterile Products |
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1. Test for Specified Micro-organisms1
(Ph. Eur. method 2.6.13)
1 INTRODUCTION
The tests described hereafter will allow determination of the abnce or limited occurrence of specified micro-organisms that may be detected under the conditions described.
The tests are designed primarily to determine whether a substance or preparation complies with an established specification for microbiological quality. When ud for such purpos, follow the instructions given below, including the number of samples to be taken, and interpret the results as stated below.
Alternative microbiological procedures, including automated methods, may be ud, provided that their equivalence to the Pharmacopoeia method has been demonstrated.
2 GENERAL PROCEDURES
The preparation of samples is carried out as described in general chapter 2.6.12.
If the product to be examined has antimicrobial activity, this is insofar as possible removed or neutralid as described in general chapter 2.6.12.
If surface-active substances are ud for sample preparation, their abnce of toxicity for micro-organisms and their compatibility with inactivators ud must be demonstrated as described in general chapter 2.6.12.
3 GROWTH-PROMOTING AND INHIBITORY PROPERTIES OF THE MEDIA, SUITABILITY OF THE TEST AND NEGATIVE CONTROLS
The ability of the test to detect micro-organisms in the prence of the product to be tested must be established. Suitability must be confirmed if a change in testing performance, or the product, which may affect the outcome of the test is introduced.
3-1 Preparation of test strains童年是一首歌
饺子的制作方法
U standardid stable suspensions of test strains or prepare them as stated below. Seed lot culture maintenance techniques (ed-lot systems) are ud so that the viable micro-organisms ud for inoculation are not more than 5 passages removed from the original master ed-lot.
3-1-1 Aerobic micro-organisms Grow each of the bacterial test strains parately in cain soya bean digest broth or on cain soya bean digest agar at 30-35 °C for 18-24 h. Grow the test strain for Candida albicans parately on Sabouraud-dextro agar or in Sabouraud-dextro broth at 20-25 °C for 2-3 days.
∙ — Staphylococcus aureus such as ATCC 6538, NCIMB 9518, CIP 4.83 or NBRC 13276;
7岁女孩∙ — Pudomonas aeruginosa芬森 such as ATCC 9027, NCIMB 8626, CIP 82.118 or NBRC 13275;
∙ — Escherichia coli such as ATCC 8739, NCIMB 8545, CIP 53.126 or NBRC 3972;
∙ 橹怎么读— Salmonella enterica subsp. enterica 操弄rovar Typhimurium, such as ATCC 14028 or, a
s an alternative, Salmonella enterica subsp. enterica rovar Abony such as NBRC 100797, NCTC 6017 or CIP 80.39;
∙ — Candida albicans such as ATCC 10231, NCPF 3179, IP 48.72 or NBRC 1594.
杜聿明简历U buffered sodium chloride-peptone solution pH 7.0 or phosphate buffer solution pH 7.2 to make test suspensions. U the suspensions within 2 h or within 24 h if stored at 2-8 °C.
3-1-2 Clostridia U Clostridium sporogenes such as ATCC 11437 (NBRC 14293, NCIMB 12343, CIP 100651) or ATCC 19404 (NCTC 532 or CIP 79.03) or NBRC 14293. Grow the clostridial test strain under anaerobic conditions in reinforced medium for clostridia at 30-35 °C for 24-48 h. As an alternative to preparing and then diluting down a fresh suspension of vegetative cells of Cl. sporogenes画皮网团购, a stable spore suspension is ud for test inoculation. The stable spore suspension may be maintained at 2-8 °C for a validated period.
3-2 Negative control
To verify testing conditions, a negative control is performed using the chon diluent in place of the test preparation. There must be no growth of micro-organisms. A negative control is also performed when testing the products as described in ction 4. A failed negative control requires an investigation.
3-3 Growth promotion and inhibitory properties of the media
Test each batch of ready-prepared medium and each batch of medium prepared either from dehydrated medium or from ingredients.
Verify suitable properties of relevant media as described in Table 2.6.13.-1.
Test for growth promoting properties, liquid media
Inoculate a portion of the appropriate medium with a small number (not more than 100 CFU) of the appropriate micro-organism. Incubate at the specified temperature for not more than the shortest period of time specified in the test. Clearly visible growth of the micro-organism comparable to that previously obtained with a previously tested and approved batch of medium occurs.
