.jpeg "Sterility Test Procedure")
1.0
Purpose:
1.1 To establish a formal procedure
for sterility test procedure of Raw Material and Finish Product.
2.0
Scope:
2.1 This
SOP applies to Microbiological Laboratory.
3.0
Responsibilities:
3.1
Officer (Microbiology) is responsible for
performing the performing the sterility test.
3.2 The Head of Quality Control/In-Charge is responsible for the compliance of this SOP.
3.3 Head of QA is responsible for SOP compliance.
4.0 Abbreviations and Definitions
4.1 QC: Quality Control
4.2
WFI: Water for
Injection
4.3
PW:
Purified Water
4.4
FTM:
Fluid Thioglycolate Medium
4.5
TSB: Tryptone
Soya Broth
5.0
Materials
and Equipment
5.1
Apparatus
and reagent:
5.1.1
Water
bath
5.1.2
Conical flask 500ml
5.1.3
Screw Cap Bottles 100ml
5.1.4
Filtration Apparatus
5.1.5
Foil for wrapping
5.1.6
Sterile Filter Paper
5.1.7
Ethanol or other disinfectants
5.1.8
Distilled water and DM water
5.1.9
Medium: Tryptone
Soy Broth
5.1.10 Medium: Fluid Thioglycolate Medium
5.1.11 Medium: Peptone Water
5.1.12 Laminar Air Flow Unit
5.1.13 Autoclave
5.1.14 Incubator
5.1.15 Microbial Culture
6.0
Precaution
/ Health and Safety Considerations
6.1
Maintain
Aseptic Condition during sterility test procedure.
7.0
Procedure:
7.1
Preparation
of Fluid Thioglycolate Medium
Media
is prepared according to SOP.
7.2
Preparation of Tryptone Soy Broth
Media is prepared according to SOP.
7.3 Preparation of Peptone Water
Media is prepared according to SOP.
7.4
Preparation
of sample:
7.4.1
Sample plan:
The sampling plan is
dependent on some factors such as quantity of sample, type of sample. Minimum
20 mg / ml sample to be tested.
7.5
Membrane Filtration Method:
7.5.1
All
Sample container surface and personnel hands surface are sterilized by
Isopropyl alcohol.
7.5.2 All glassware, membrane filter apparatus
are sterilized by autoclave.
7.5.3 Sample is taken according to batch size
or quantity as compendial (BP/USP) recommendation.
7.5.4 Pre-sterilize filter paper are set on
the filter paper apparatus holder.
7.5.5 Add sample into the peptone water and
mix properly.
7.5.6 Prepared sample is directly poured into
the filter unit.
7.5.7 Then Vacuum
pump is ‘Switched ON’ for applying negative pressure.
7.5.8 After that
Vacuum pump is Switched ‘OFF’.
7.5.9 Then filter paper cup is removed from filter
holder apparatus and collected the filter paper.
7.5.10 Filter paper
then cut into two equal pieces aseptically by sterile scissors.
7.5.11 One piece put
into FTM, another one in TSB.
7.5.12 Prepare a set
of positive control of FTM and TSB.
7.5.13 One FTM and
one TSB maintain as negative control without product and culture inoculation.
7.5.14 Incubated the
FTM at 30-35°c for 14 days for anaerobic and aerobic bacteria, TSB at 20-25°c
for 14 days for fungi and aerobic bacteria.
7.6
Direct Transfer Sterility Testing
7.6.1
All
Sample container surface and personnel hands surface are sterilized by Isopropyl
alcohol.
7.6.2
All
glassware are sterilized by autoclave.
7.6.3
Sample
is taken according to quantity as compendial (BP/USP) recommendation.
7.6.4
Approximately
20 mg / ml sample is transferred to a conical flask containing peptone water
and mix well.
7.6.5
From
this conical flask, add 10 ml of sample to FTM medium and add 10 ml of Sample
to TSB medium.
7.6.6
Prepare
a set of positive control of FTM and TSB.
7.6.7
One
FTM and one TSB maintain as negative control without product and Culture
inoculation.
7.6.8
Incubate
the FTM at 30-35°c for 14 days for anaerobic and aerobic bacteria, TSB at
20-25°c for 14 days for fungi and aerobic bacteria.
7.7
Specification:
7.7.1
No
turbidity form within 14 days of sample inoculated to FTM and TSB.
7.8
Method:
7.8.1
Based
on turbidity form method in the liquid broth medium from which bacteria and
fungi grow under aerobic conditions.
8.0 Reference
Document
8.1
USP <71>
9.0 Annexure
9.1
Annexure-I: Sterility Test Report
10.0 Revision History
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Revision
No. |
Brief reason for the revision |
Effective Date |
Remarks |
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01 |
New |
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11.0
Training
11.1
Head of Quality Control or his/her
nominee shall give the SOP training before effective date.
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Name
of the Product/Material: |
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Batch No.: |
Batch Size/Quantity: |
QC Ref. No.: |
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Mfg. Date: |
Exp. Date: |
Tested Quantity: |
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Origin/ Source: |
Test Starting Date: |
Test Completion Date: |
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Test Condition: Fluid Thioglycolate Media at 30-350C
for 14 days; Trypton Soya Broth Media at 20- 250C for 14 days.
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Observation: |
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Observed day |
Fluid Thioglycolate
Media |
Trypton
Soya Broth Media
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(Test Sample) |
(Negative
Control) |
(Positive
control) |
(Test Sample) |
(Negative
Control) |
(Positive Control) |
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Day -4 |
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Day- 5 |
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Day -6 |
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Day -7 |
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Day- 8
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Day- 9
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Day -10 |
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Day -11 |
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Day -12 |
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Day -13 |
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Day -14
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Comments
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Tested by: Executive / Sr. Executive, Microbiology |
Checked by: Executive / Sr. Executive, QC |
Approved by: Head of QC / QA |
q PASSED q REJECTED |
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Signature & Date |
Signature & Date |
Signature &
Date |
Legends: (+)
Presence of growth, (–) Absence of growth.
