MICLAB 060 Sample

Standard Operating Procedure Title: Micro Laboratory Procedure for Sterility Testing ______________________________________________________________________________________

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6. Repeat Testing of Sterile Products..................................................................................................... 7 7. Media Check after Sterility Testing (Stasis Test) ................................................................................ 8 8. Validation and Revalidation of Sterility Test Methods ......................................................................... 8 9. Appendix 1 - Flowchart of Sterility Test results - Interpretation and Retests.......................................11 10. Summary of Changes.......................................................................................................................12

Procedure

1. Obtaining of Samples

1.1. Sampling Production personnel conduct all sampling in the following areas. Details of these sampling procedures are outlined in MICLAB 095. Terminally Sterilised Products Samples are to be selected randomly from throughout the autoclave load.

Aseptically Filled Products Samples are to be selected from the beginning, middle and end of each batch, plus the first units filled after any prolonged downtime (greater than one hour). Samples are to be of saleable standard, not rejects.

1.2. Sample Size required for Initial Sterility Testing Determine the sample size of the product type and batch size.

2. Setting up a Testing Session

2.1. Checking the testing requirements for each product code Every product sampled must be documented properly in details when setting up a test session. This will indicate what tests are required for each product code. Always cross-check the BPN on the sample container against the sample ID sheet.

2.1.1. Parametric Release Not all terminally sterilised products require sterility testing. Check for Parametric Release status of any product code.

2.1.2. Sterility Test These products require sterility testing. A standard sterility testing session is comprised of 4 products and a sterile control to be membrane filtered and any number of direct inoculations along with a suitable sterile control.

2.1.3. Bacterial Endotoxin Check if a product requires Bacterial Endotoxin testing and also by which test method, either gel clot or KCA. If the product requires Bacterial Endotoxin testing, separate these samples from the sterility samples. Ensure that these samples are labelled with the Product Name & strength, batch, code and place these in the receptacle for the type of Endotoxin test required.

2.1.4. Biological Assay If a biological assay is required separate the samples from the Sterility samples. Ensure these samples are labelled with the Product Name, batch and code and are given or communicated to the technician responsible for Assay testing.

2.2. Known Sterile Controls




Incubate the TSB and FTM bottles as described below in point 3.6 Incubation. Secondary Transfer Process After not less than 14 days of incubation and working within the sterility test LAF, aseptically transfer the following: - 1 mL of the initial FTM media into a new bottle of 100mL FTM, and, - 1 mL of the initial TSB media into a new bottle of 100 mL TSB. Continue the incubation of the initial media and transfer media bottles as follows:

FTM Not less than 4 days @ 322C. TSB Not less than 4 days @ 222C. Examine the media for growth at intervals during the incubation period. If no evidence of growth is found the batch meets the requirements for the Test of Sterility.

3.2. Agar Plates One irradiated Letheen Agar plate and one irradiated Nutrient Agar plate are to be taken into the Sterility Test room for every sterility test session. The Nutrient Agar plate is to be left exposed on the Laminar Flow bench (sterile Environmental) throughout the entire time of the sterility test session. The Letheen Agar plate is to be used to record the Operator's finger print impressions during the test session or at its conclusion. If a needle stick injury occurs, before leaving the sterility test room to change gloves, conduct a finger print impression. Use an additional Letheen Agar plate to monitor the finger impressions at the conclusion of the session. Four (4) contact plates are used for personnel monitoring of the hood, sleeve, chest and leg after all testing has been completed, on exiting the change room. NOTE: If a double consecutive session is being tested, perform personnel monitoring after the second session only. Record the Name, Date and Session details on the plates. Incubate all plates according to MICLAB 045. Record results in the Monitoring Results for Sterility Test Sessions Finger, Fallout, Environmental and Personnel Results File. NOTE: After recording the finger print impression, the Operator must don a new pair of sterile gloves in the change room and disinfect their hands with 70% IPA prior to resumption of testing. See MICLAB 045 for limits.

3.3. Environmental Monitoring Environmental monitoring must be performed with each Sterility Testing Session and in each room and transfer hatch in use, on a daily basis. See the Table below for details:

Area Environmental Monitoring Required Frequency Laminar Flow Cabinet 1000L Air Sample

1 x Contact Plate Each Session Each Session

Change Room 200L Air Sample 1 x Contact Plate (Floor)

Daily, when in use Daily, when in use

Transfer Hatch 200L Air Sample 1 x Contact Plate

Daily, when in use Daily, when in use

Sterility test Room 200L Air Sample 1 x Contact Plate (Floor) 1x Contact Plate (Trolley)

Daily, when in use Daily, when in use Daily, when in use

The environmental Agar plates and contact plates are to be labelled with all details and incubated according to MICLAB 045. Results from the Laminar Flow monitoring are to be entered in the Monitoring Results for Sterility Test Sessions Finger, Fallout, Environmental and Personnel Results File. Results from ALL environmental samples are to be entered in the Environmental Results Air / Surfaces Non-Sterile File. After the total count is recorded,




The product is to be regarded as a separate session with a known sterile control. No other product is to be tested during this procedure. a) Non-injectable, Non-filterable products

For non-injectable, non-filterable products follow the same procedure as for the initial test. The known Sterile Control is Paraffin Oil in this test session.

b) Aqueous Solutions 1. For aqueous solutions the same quantity of product is filtered. 2. Include a known sterile control of X units of Water for Injection that has been

autoclaved at least twice in the test session. 3. Wash the canister filters with 3 x 100 mL of Sterile Peptone Water. These 3

washes should be individual. Fill the canisters with the appropriate medium, seal and incubate as for an initial sterility test.

6.4. Recording of the session and results Details of the test sessions and the results are to be recorded in the log book.

6.5. Interpretation of sterility repeat test results

6.5.1. If no evidence of microbial growth is found in the repeat test, the product examined complies with the test for sterility. If microbial growth is found in the repeat test, the product examined does not comply with the test for sterility.

7. Media Check after Sterility Testing (Stasis Test)

7.1. Membrane Filtration and Direct Inoculation Once every 12 months media containers from a product tested by Membrane Filtration and also if possible those from a product tested by Direct Inoculation must be subjected to a challenge by a low number of microorganisms after the sterility test has been completed. This is to ensure that any inhibitory effects present in the products have been successfully inactivated by the dilution effect of the testing procedure, or the washing procedure as appropriate to the Test Method, and hence any microorganisms that may have been present in the product would have been able to grow in the media when the product was tested. It is important that on an annual basis, the range of sterile products are to be subjected to a Stasis Test. A Stasis Check on Sterility Tests Form must be filled out on an annual basis detailing the product type and date the Stasis test was conducted. NOTE: The product types listed are not exhaustive; they represent the range of products manufactured by the company and list only the specific products most likely to be inhibitory to the Sterility test.

7.2. See MICLAB 090 for the procedure of Stasis tests and to determine the method of preparation of Stock Suspensions of micro organisms used for Stasis work and media checks.

8. Validation and Revalidation of Sterility Test Methods

8.1. General Requirements 8.1.1 All products, which require sterility testing, must have a corresponding validated

method. 8.1.2 Validation is required for each new product formulation, changes in product

formulation, changes in containers / closure types and any other changes that impact the status of a validated system, process or equipment.

8.1.3 Re-validations of existing methods are to be performed once every 5 years. 8.1.4 A revalidation of the sterility test system, using at least one product is to be

conducted on an annual basis.




9. Appendix 1 - Flowchart of Sterility Test results - Interpretation and Retests

Initial Test

Test Result = no growth

Test Result = Growth

Control Result = growth

Control Result = no growth

PASS FAIL

Repeat test

Test result = no growth

Test Result = Growth

Problem with session?

YES NO

Does the organism match the

environment?

YES

NO

Form 680 Issue date:

Sterility Test Failure Investigation (Ref. MICLAB 060)



Table Contents DR____________ 1. Purpose of report 2. Batch Details 3. Batch Manufacturing review and Sterility testing 4. Summary of Conclusions found 5. Possible causes 6. Corrective action 7. Preventative action 8. Disposition of filling room 9. Documentation Approval of Investigation

1 Purpose of report The purpose of this document is to provide a summary report of the incident investigation conducted into: 2 Batch Details Product Name Batch # Date of Manufacturing Filling Room # Product Code Filling Machine Container and size Terminally sterilized Y/N Autoclave Cycle Sterility Test Details Date Technician

Test room Number of products

tested

Time of test session Session number that day

Validated test method Tested to Specification

Date of results Test session results

Contaminant/s type Negative control result

3 Batch review from manufacturing and testing. Product Manufacturing Review REVIEW AREA DETAILS & Results REVIEWED BY Dispensing results: Review of raw materials used for the manufacture

Batch # Materials code Validated test method Results of tests

Sign: Date: Review Attached: Yes/No

Dispensing of the raw materials


Sterilization cycles for components

Autoclave Cycle# Date Result






Interview of Operator for any excursion of procedures:

Name: Date


Training records Gowning validations

Name Training Record


Operators Finger Dabs

Name Results


Operators Uniform

Name Results


Packing Line:

Autoclave cycle finished product

Autoclave # Cycle # Result


Issues on Inspection / Finishing line


QA Review Manufacturing: Product Review of past 12 months of results


Deviation Reports Review


Similar incidents Sign:

Date: Review Attached: Yes/No

Change control history review


Audit of security card swipe


Review of Media run reports past 12 months


Summary Micro. Lab. to place a HOLD on adjacent & subsequent batches until establishing a cause which will eliminate their risk of contamination and also; Batches on HOLD (Q) Batch # Code Product & % Container Size Sterility Testing Review REVIEW AREA DETAILS REVIEWED BY Results of known sterile controls


Level of false positives in routine testing (previous 12 months)


Level false positives in known sterile controls previous 12 months)


Review of sampling procedures


Date done: Notify QA Manager





Interview of Technician


Sterility Log Book


Comparison of Isolates Date of Streaking all Isolates:_________________ Technician:________________________________ Source of Isolate

No. of Colonies

Colony Morphology

Microscopic Appearance

Product

N/A

Finger Dab Plate

Uniform Plate

Laminar Flow Contact plate

Laminar Flow Air Sample

Fallout Plate

IPA Exp. Date: Bucket 1

N/A

Bucket 2

N/A

4. Summary of Conclusions found 5. Possible cause/ Root Cause

MICLAB 060 Sample

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