gpt protocol - trypcase soy agar

Protocol for Growth Promotion TestTrypcase Soy Agar Plates (Tryptone Soya Agar)

Document No. Effective dateVersion No. Page No. of 8

Prepared byName:

Sign & DateApproved by:

Name:

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I. Objective:

Growth promotion test is performed on microbiological culture media to assess the ability of the media to sustain growth. This protocol outlines the procedure for Growth promotion testing at SPT. The micro-organisms used are recommended by the European Pharmacopoeia & USP.

II. Scope:

The scope of this protocol is applicable to the Growth Promotion test, certification, usage and reconciliation of Trypcase Soy Agar (TSA) plate received at MICRO lab.

III. Procedure:

Once a new lot of media from an approved vendor is received, it is necessary to certify the lot. The lot may be certified by performing growth promotion tests.

Documentation of growth promotion tests will be done in this document including corresponding certificate of analysis and usage and reconciliation. Each lot will have separate protocol / report. The report will be completed and archived on exhaustion of specific lot of the media.

Growth promotion test is performed by comparing one plate of new lot of media to two plates of previously certified lot. A certified lot is a lot of agar of known fertility. In-case of initial lot, this shall not be applicable. The colony forming unit (CFU) count in the new lot of media is then compared to the mean count of the certified plates and the percentage recovery is calculated.

A. Determination of microorganisms that are needed to test the medium:

Medium Organism Incubation temperature

Incubation time

Trypcase soy Agar (Tryptone Soya Agar)

Staphylococcus aureus ATCC 6538 - represent as gram positiveEscherichia coli ATCC 8739 - represent as gram negativeCandida Albicans ATCC 10231 - represent as yeast

30oC to 35oC

30oC to 35oC

30oC to 35oC

≤ 3 days



IV. Receipt, verification and storage of TSA Plates:

Name of the product (as mentioned on Certificate, PO and product label)

Supplier Catalogue No.

Batch No.

Mfg. Date

Exp. Date

Quantity received

Purchase Order reference number & date

Supplier

Storage condition

CoA availability

Pack integrity check Satisfactory / Not satisfactory

Details of damage / remarks (if any):

Received and verified by: Name:

Signature and date:

Final remarks of consignment Accepted / Rejected

On satisfactory verification, apply quarantine label on the pack and store at respective storage conditions / location.

V. Sampling Instruction:Two plates to be tested for growth promotion tests. Two plate to be used a negative control.



Prepared byName:


Name:

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Preparation of culture:

Organism Lot No.Escherichia coli ATCC 8739Candida Albicans ATCC 10231Staphylococcus aureus ATCC 6538

DescriptionEscherichia coli ATCC 8739 Lot No.: Candida Albicans ATCC 10231 Staphylococcus aureus ATCC 6538Date & time of

preparationPerformed by (Sign & date)

Date & time of preparation

Performed by (Sign & date)



Remove the hydrating fluid vial and vial of lyophilized strain preparation from refrigerated storage. Allow unopened vial of lyophilized pellets to equilibrate to room temperatureWhile pellets are equilibrating, pre-warm the hydrating fluid to 35oC (at least 30 minutes)With a sterile forceps, transfer two (2) pellets into



DescriptionEscherichia coli ATCC 8739 Lot No.: Candida Albicans ATCC 10231 Staphylococcus aureus ATCC 6538Date & time of

preparationPerformed by (Sign & date)





the 2 ml vial of hydrating fluid. (do not remove the desiccant from vial). Two pellets must be used to obtain the challenge concentration of <100 CFU per 0.1 ml. Immediately stopper and recap the pellet vial and return the remaining lyophilized material to refrigerated storage of 2oC to 8oCImmediately re-cap the vial with the hydrated material and place into a 34oC to 38oC incubator for thirty (30) minutes to assure complete hydrationImmediately following incubation vortex the hydrated material to achieve a homogeneous suspension



VI. Testing Procedure:

DescriptionSpecification /

procedureObservation

Done by (Sign & date)

Escherichia coli ATCC 8739 Lot No.:

Candida Albicans ATCC 10231

Staphylococcus aureus ATCC 6538

Date of testing (start date):Date completed (after observation of results):

Label plates with the organism name, duplicate number, date of inoculation and media type

To comply

Using sterile pipette, pipette-out 0.1 ml of each microorganism solution to the media plates

To comply Volume transferred:

Lot No. of sterile Pipette:

Volume transferred:


Volume transferred:


Using sterile spreader spread the solution evenly across the surface of the agarAllow plates to dry before inverting and incubation.

To comply Lot no. of sterile spreader:

Lot no. of sterile spreader:

Lot no. of sterile spreader:

Two agar plate (of new lot) incubated along with the test plates without any inoculation. This will act as negative control

To comply

Record the time and date of incubation period in the Growth Promotion Test Report of individual medium.

To comply Incubator No.:

Date Incubated:

Time Incubated:

Incubator No.:

Date Incubated:

Time Incubated:

Incubator No.:

Date Incubated:

Time Incubated:



DescriptionSpecification /

procedureObservation

Done by (Sign & date)

Escherichia coli ATCC 8739 Lot No.:



After incubation, remove all plates from the incubator, record the time and date that the plates removed.

To comply Date removed:

Time removed:

Date removed:

Time removed:

Date removed:

Time removed:

Inspect each plate for growth, counting all CFU present on each plate, The percentage recovery is calculated using the formula given below:

For New Lot

Average Test count (new lot) (CFU)% recovery = ----------------------------------------- X 100

Average count of approved plates

The average count must be between 10 to 100 CFU.

% recovery must not differ by a factor greater than 2 (i.e., 50% to 200%)

No growth must be observed in negative control

New lot:Plate 1:

Plate 2

Average:

Previously approved lot:Plate 1:

Plate 2:

Average:

% recovery:

Negative control observation:

New lot:Plate 1:

Plate 2

Average:


Plate 2:

Average:

% recovery:


New lot:Plate 1:

Plate 2

Average:


Plate 2:

Average:

% recovery:




Prepared byName:


Name:

Sign & Date

VII. Certificate of Analysis

Name of the Media Delivery Note No.

Batch No. / Lot No. Quantity received

Mfg Date Quantity approved

Expiry date Quantity rejected

Date of receipt Date of Certification

OrganismSpecification / results

Remarks (Pass / Fail)Average test count

% recovery

Escherichia coli ATCC 8739



Review remarks of supplier CoATo confirm the purity and authenticity of supplied culture

Acceptance Criteria: Average test count: Not More Than 100 CFU; test plate count must not differ from control plate count by factor greater than 2

Remarks: The received consignment Complies / Does not complies to the specifications. The material is approved / rejected for further intended use

Prepared by (Signature and date) Approved by (Signature and date)



VIII. Usage log: Stock control and reconciliation (use additional sheets where needed)

Quantity Released

Product Name / Item B. No. Test Name Date used Quantity used

Remarks:

Reconciliation performed by:

Name:

Signature & Date

Verified by:

Name:

Signature & Date

gpt protocol - trypcase soy agar

Documents

lot of agar

certified lot

sign date protocol

plate of new lot of

date quantity

certified plates

exhaustion of specific

case of initial lot