Media Shelf Life Validation Protocol | USP & Compliant.
| PHARMA MICRO HUB | VALIDATION PROTOCOL | |
| TITLE: SHELF LIFE DETERMINATION OF PREPARED MICROBIOLOGICAL MEDIA | ||
| Protocol No: PMH/VP/001-00 | Effective Date: April 2026 | |
1.0 OBJECTIVE
To define the detailed procedure for determining the shelf life of prepared microbiological media by evaluating pH stability, physical integrity, and growth promotion capability over a 14-day study period.
2.0 SCOPE
This protocol is applicable to all dehydrated media lots prepared, sterilized, and stored at 22.5 ± 2.5°C within the Microbiology Laboratory of Pharma Micro Hub.
3.0 RESPONSIBILITY
| Designation | Responsibility |
|---|---|
| Microbiologist | Execution of media preparation, sterilization, pH measurement, physical checks, and GPT. |
| Head – Microbiology | Technical review of raw data, calculation of recovery percentages, and report verification. |
| Head – QA | Final approval of the protocol and authorization of the established shelf life. |
4.0 PRE-REQUISITES
- Validated Autoclave (Cycle: 121°C for 15 minutes).
- Calibrated pH Meter with temperature compensation.
- Standardized ATCC Microbial Cultures (e.g., B. subtilis, S. aureus).
- Laminor Air Flow chamber
5.0 PROCEDURE
5.1 Media Preparation: Reconstitute dehydrated media powder in purified water and mix properly.
Sterilize at 121°C for 15 minutes at validated autoclave as per SOP.
Cool to 45-50°C and pour 20-25ml into sterile plates aseptically under LAF.
5.2 Physical Check: Visually inspect the plates for colour change, excessive moisture, cracks or microbial contamination.
5.3 pH Procedure: Calibrate the pH meter using standard buffers then Measure pH of solidified agar and broth media at 25°C.
Acceptable limit is ±0.2 or as per manufacture recomenadtions.
5.4 GPT Performance: Inoculate test organisms in media with 10-100 CFU of challenge organisms. Incubate per USP <61>/<62> and confirm recovery is 50-200% compared to fresh control media.
6.0 ANNEXURES
Annexure I: Media Preparation & GPT Log.
Annexure II: Physical & pH Observation Sheet.
7.0 REFERENCES
- USP <61>: Microbial Enumeration Tests.
- USP <62>: Tests for Specified Microorganisms.
8.0 ABBREVIATIONS
GPT: Growth Promotion Test
LAF: Laminar Air Flow
USP: United States Pharmacopeia
CFU: Colony Forming Unit.
| PHARMA MICRO HUB | VALIDATION SUMMARY REPORT | Report No: PMH/VR/001-00 |
| REF PROTOCOL NO: PMH/VP/001-00 | Date: April 2026 |
1.0 OBJECTIVE
To summarize and evaluate the analytical data obtained during the 14-day hold-time study to establish the final shelf life for prepared media.
2.0 SCOPE
This report covers the stability evaluation of media batches stored under controlled laboratory conditions (22.5 ± 2.5°C).
3.0 RESPONSIBILITY
| Designation | Responsibility |
|---|---|
| Microbiology Team | Compiling results from Day 0, Day 7, and Day 14 into the summary report. |
| QA Department | Reviewing summary against protocol and authorizing the final shelf life. |
4.0 REQUISITES USED
Qualified ATCC organisms, validated dehydrated media lots, and calibrated pH analysis systems.
5.0 PROCEDURE FOLLOWED
The study was executed per Protocol PMH/VP/001-00. Testing for GPT, pH, and physical appearance was performed at Day 0, Day 7, and Day 14 intervals.
6.0 RESULTS SUMMARY
| Media Name | Parameter | Day 0 | Day 7 | Day 14 |
|---|---|---|---|---|
| SCDA / SCDM | pH & Appearance | Pass | Pass | Pass |
| SDA | GPT Recovery | Pass | Pass | Pass |
7.0 CONCLUSION
All prepared media batches met the acceptance criteria for pH stability, physical integrity, and growth promotion capability (50-200% recovery) throughout the 14-day study duration.
8.0 RECOMMENDATIONS
The validated shelf life for prepared microbiological media is established as 10 days from sterilization date at 22.5 ± 2.5°c.
ANNEXURES
Annexure I: GPT Recovery Log Template
| Organism | Control (CFU) | Test (CFU) | % Recovery |
|---|---|---|---|
Annexure II: pH & Physical Appearance Log
| Interval | pH Observed | Physical Appearance |
|---|---|---|
| Day 0 / 7 / 14 |
