Site icon Pharma Dekho

sop for preservatives efficacy test

 

sop for preservatives efficacy test

 

1.0 OBJECTIVE 

1.1 To lay down a procedure for testing of Efficacy of Preservatives.

2.0 SCOPE

2.1 This SOP is applicable for testing of Efficacy of Preservatives added in products

3.0 RESPONSIBILITY

3.1 Microbiologist QC

4.0 ACCOUNTABILITY



4.1 QC Manager

5.0 PROCEDURE

5.1 Ensure the all the media employed should be tested for their growth promoting properties.
5.2
Inoculum Preparation:
5.2.1 Inoculate the surface of the Soybean casein Digest Agar and Sabouraud Dextrose Agar media plates / slants with 1 ml of the microorganisms from the recent culture suspension as given in Table-1.
5.2.2 Incubate the plates/ Slants at suitable conditions as given in the Table-1.

5.2.3 Harvest the cells except Aspergillus niger after the incubation by washing the growth on the surface of the respective solid agar media using Sterile Normal Saline (0.9%w/v), collect it in a sterile container and treat this as Stock solution.
5.2.4
5.2.5

For harvesting of Aspergillus Niger, Use Normal saline with 0.05% Polysorbate 80.
From the Stock solution, do tenfold serial dilutions using sterile normal Saline as diluent to get a count of 1 x 108 CFU / ml.
5.2.6 Determine the number of CFU per ml of these harvested suspensions by plate count Method using suitable Agar media and incubate at appropriate temperatures.
5.2.7 Use the initial count of suspensions to calibrate the size of inoculum to be used in the Test.
5.2.8 Use the Bacterial and Candida albicans suspensions within 24 hours of harvest.

5.2.9 Fungal suspensions shall be refrigerated and used up to 7 Days.

5.2.10 For microorganisms and media to be used, incubation conditions, microbial recovery time, refer to Table-1 given below.
Table-1
Organism Suitable Medium Incubation
Temperature Inoculum
Incubation Time Microbial Recovery
Incubation Time
Escherichia coli
(ATCC No. 8739) Soybean–Casein Digest Broth;
Soybean–Casein Digest Agar 32.5 ± 2.5 18 to 24 hours 3 to 5 days
Pseudomonas aeruginosa
(ATCC No. 9027) Soybean–Casein Digest Broth;
Soybean–Casein Digest Agar 32.5 ± 2.5 18 to 24 hours 3 to 5 days
Staphylococcus aureus
(ATCC No. 6538) Soybean–Casein Digest Broth;
Soybean–Casein Digest Agar 32.5 ± 2.5 18 to 24 hours 3 to 5 days
Candida albicans
(ATCC No. 10231) Sabouraud Dextrose Agar;
Sabouraud Dextrose Broth 22.5 ± 2.5 44 to 52 hours 3 to 5 days
Aspergillus niger
(ATCC No. 16404) Sabouraud Dextrose Agar;
Sabouraud Dextrose Broth 22.5 ± 2.5 6 to 10 days 3 to 7 days



5.3 Test Procedure:
5.3.1 Use five Product containers if the volume per container is sufficient.
5.3.2 If the volume is not sufficient, transfer the product into five sterile screw capped containers.
5.3.3 Reconstitute the product container if it is in dry powder form, as per the instruction on the label.
5.3.4 Standardize the volume of the inoculum to be between 0.5% and 1.0% of the volume of the product and the concentration to be between1 × 105 and 1 × 106 cfu per mL of the product.
5.3.5 Add the inoculum into the product containers using Sterile Syringe or Pipette.
5.3.6 Determine the initial count of the inoculated containers by Plate Count Method.
5.3.7 Incubate the inoculated containers at temperature 22.5C ± 2.5
5.3.8 Withdraw samples from containers at the intervals of 14 and 28 Days and determine the number of CFUs present in Sample containers by the Plate Count Method.
5.3.9 Observe and record any changes in appearance of the sample during the sampling as per Annexure-I.
5.3.10 Incorporate an Inactivator in the diluents such as sterile normal saline (0.9 % w/v) with polysorbate-80 (0.05 % w/v) and 0.05 % Soya lecithin or with peptone (0.1 % w/v) and 0.05 % Soya lecithin or sterile normal saline (0.9 % w/v) with 0.05 % polysorbate-80.
5.3.11 Using the Initial count of the test preparation, calculate the change in log10 values of the concentration of cfu per mL for each microorganism at the respective intervals.
5.3.12 Express this in terms of Log Reduction as per Annexure-I and report as Complies if results are complying to the criteria as given in Table-2.



Table –2
S.No. Micro organism Acceptance Criteria
1 Bacteria Not less than 1.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days.
2 Yeast and Molds No increase from the initial calculated count at 14 and 28 days.
Note: No increase is defined as not more than 0.5 log10 unit higher than the previous value Measured.

6.0 ABBREVIATIONS

Sr. No. Abbreviation used Extended Form
1.0 SOP Standard Operating Procedure
2.0 QC Quality Control

7.0 ATTACHMENTS (ANNEXURE)

Annexure –I : Preservative Efficacy Test report

8.0 REFERENCE

 

Sr. No. Reference Title
1 In House

 

Annexure –I

PRESERVATIVE EFFICACY TEST REPORT

Name of the Product
Batch No.
A.R.No
Date of analysis
Date of report

Media Details:

 

Day of Analysis 1st Day 14th day 28th day
Media used
Lot No.:  

 

 

Autoclave Lot No.:  

 

 

Incubation Period

 

1st  Day 14th  day 28th  day
Incubation

Started at

/ Date

Bacterial Cultures

Incubator No:

(30ºC – 35ºC)

Fungal Cultures

Incubator No:

(20ºC – 25ºC)

Incubation

Completed at / Date

Bacterial Cultures

Incubator No:

(30ºC – 35ºC)

Fungal Cultures

Incubator No:

(20ºC – 25ºC)

Incubation Details: 

 

Name of the Microorganism Day of Analysis Dilution Considered in the Calculation Test Count  

% recovery

Final Count*   x 100

Initial Count*

Escherichia coli

ATCC 8739

1st  Day
14th  Day
28th  Day
Pseudomonas aeruginosa ATCC 9027 1st  Day
14th  Day
28th  Day
Staphylococcus aureus

ATCC 6538

1st  Day
14th  Day
28th  Day
Candida albicans

ATCC 10231

1st  Day
14th  Day
28th  Day
Aspergillus niger ATCC 10231 1st  Day
14th  Day
28th  Day
Day of Sample withdrawal Appearance of the Product:
1st  Day
14th  Day
28th  Day

* Initial Count denotes the Count of 1st Day Analysis and Final Count indicates 14th  and 28th  Day Analysis.

                                                    Acceptance Criteria
Bacteria Not less than 1.0 log reduction from the initial count at 14 days, and no increase from the 14 days’ count at 28 days.
Yeast and Molds No increase from the initial calculated count at 14 and 28 days.

 

Conclusion: The preservative added is effective/ not effective and the product complies/does not comply.

 

Observed by                                                                                    Reviewed by                                                                                  Approved by

sop for calibration and validation of micro autoclave

sop for Sterility failure investigation

cleaning and operation of discard autoclave

sop for operation of fogger machine

sop for Biological assay of lactic acid bacillus

sop for preparation of culture inoculum

STP for sterility testing of sterile gloves

sop for Operation and calibration of active air sampler

sop for transfer of material for testing and sampling in sterile area

entry & exit procedure in microbiology laboratory

Growth Promotion Test In Microbiology Laboratory

Operation of B.O.D in Microbiology Laboratory

Operation of Horizontal Laminar Air Flow in the microbiology laboratory

Operation and cleaning of Pass Box.

Operation and cleaning of air sampler

Cleaning and Sterilization of Glassware

Analysis of water for microbial load in microbiology lab

Operation and temperature monitoring of Refrigerator

Fumigation of Microbiology Laboratory.

Entry & Exit procedure In Sterility Area

SOP for Microbial analysis of Raw Material Finished Products

SOP for Operation & Calibration of pH meter in Micro Department

SOP for Operation & Calibration of pH meter in Micro Department

SOP Operation and calibration of Hot Air Oven In Microbiology

SOP for operation cleaning & calibration of Digital colony counter

SOP for Operation And Cleaning of Microscope

sop for Media Preparation and Consumption

sop for Receipt Storage and Usage of Culture Media

sop for Cleaning Sanitization And Disinfection In Microbiology

sop for Environmental monitoring of all the Classified area

sop for Handling and Sub culturing of Microbial cultures

sop for Media Growth Promotion Test and various Microbiological test

sop for BOD incubator operation and cleaning

sop sampling of water for microbiological analysis

sop for Disinfectant Efficacy Test

sop for for cleaning and operation of vortex mixture

sop for Temperature & Relative Humidity Monitoring

sop for Operation and Calibration of Heating Block

sop for Sterility Testing of Microbiology

sop for Disposal of Culture Media

sop for Drain point of Microbiology

sop for entry & exit procedure In Microbial limit test and Biosafety

sop for Gram Staining of Bacteria in Microbiology Laboratory

sop for Monitoring of Compressed Air/gases for microbiological analysis

sop for BET (Bacterial Endotoxin) test in Microbiology

sop for receipt storage and Determining the population of Biological indicators

sop for qualification of analyst microbiologist

sop for Bioburden test of Packing materials in Microbiology Laboratory

sop for microbiological assay of erythromycin antibiotic

sop for liquid particle counter

sop for operation and calibration of digital zone reader

sop for monitoring of ultraviolet efficiency LAF and pass box

microbiological assay of cyanocobalamin or vitamin B12

gowning procedure for microbiological testing area

swab testing of various surfaces for bioburden determination

sop for endotoxin challenge test

Hold time study protocol for sterilized media

sop for personnel Qualification protocol for aseptic area

sop for sampling and testing of drain water

Sop for Operation of Airborne Particle Counter

sop for Inoculum Preparation

sop for Validation protocol of steam sterilizer autoclave

sop for pathogen detection from drain point

Sop for Analysis of Raw water Purified water water for injection and pure steam water

sop for preservatives efficacy test

sop for collection and preservation of in house isolated microorganisms

sop for Operation Calibration and Maintenance of Micropipette

Sop for UV Efficacy Test

sop for gram staining

Sop for swab testing

sop for microbiological testing of water

PROCEDURE FOR FUMIGATION

sop for depyrogenation of apparatus

sop for media preparation

sop for fertility test growth promotion test of media

sop for Operation and cleaning of moist heat sterilizer

sop for monitoring by active air sampler

sop for swab sampling and testing for clean rooms in production area

sop for monitoring in microbiology laboratory

sop for Fumigation of aseptic area and microbiology lab

sop for monitoring of personnel in aseptic area

sop for maintenance of cultures

sop for Operation and cleaning of laminar bench

preparation of settle plates

sop for monitoring of pure steam

sop for entry and exit procedure to m.l.t and b.e.t room

sop for storage of and use of media

sop for disposal of microbiological media and cleaning of microbiological glassware

 

 

 

Exit mobile version