activities carried out during the aseptic filling of vials, ampoules, and prefilled syringes in ISO Class 5 (Grade A) and surrounding ISO Class 7 (Grade B) cleanroom environments.

3. Responsibilities

• Microbiology Analyst: To perform routine microbial sampling as per protocol.
• Manufacturing Operator: To cooperate during monitoring and prevent contamination.
• IPQA Officer: To oversee sampling integrity and environment conditions.
• QC Microbiology Supervisor: To ensure compliance with testing schedules and review results.

4. Accountability

The Head of Quality Control is accountable for ensuring that microbial monitoring activities are carried out effectively, deviations are investigated, and results are recorded and reported in a timely manner.

5. Procedure

5.1 Preparation

1. Verify calibration and cleanliness status of environmental monitoring instruments (e.g., air samplers, settle plates, contact plates).
2. Ensure media used (TSA plates, SDA plates) are within expiry and approved for use.
3. Label all media with batch number, sample point ID, date, and time before sampling.
4. Ensure sampling is performed by qualified microbiologists wearing sterile gowning.

5.2 Active Air Sampling

1. Use a calibrated air sampler positioned in critical areas (e.g., near filling heads, stopper bowls).
2. Set sample volume (usually 1000 L for Grade A areas) and run per standard protocol.
3. Record data in the Environmental Monitoring Log (Annexure-1).

5.3 Passive Air Sampling (Settle Plates)

1. Place TSA plates exposed for 4 hours in Grade A and B areas, covering representative zones (e.g., filling line, operator working bench).
2. Avoid placing plates in areas with strong airflow or vibrations.
3. Retrieve plates using aseptic technique and transfer to incubation area.

5.4 Surface Monitoring (Contact Plates)

1. Monitor critical surfaces like:
   • Gloved hands
   • Filling machine parts
   • Work surfaces
   • Container closure contact areas
2. Press contact plate gently for 10 seconds without movement.
3. Replace lids and label with details.
4. Document surface locations sampled (Annexure-2).

5.5 Personnel Monitoring

1. Sample personnel gloves, sleeves, and chest area at the end of filling operation.
2. Any personnel entering Grade A zones must be monitored at least once per shift.
3. Record findings under Annexure-3.

5.6 Incubation and Result Interpretation

1. Incubate TSA plates at 30-35°C for 48–72 hours and then at 20–25°C for an additional 72 hours.
2. SDA plates used for fungal detection should be incubated at 20-25°C for 5–7 days.
3. Count colony-forming units (CFUs) and compare to alert and action limits as defined by the site’s Environmental Monitoring Program.
4. Record all observations and deviations in Annexure-4.

5.7 Action on Out-of-Limit Results

1. If action limits are breached:
   • Immediately notify QA and halt aseptic operation if applicable.
   • Initiate deviation report and root cause investigation.
   • Repeat sampling and increase frequency if required.
2. Record incident in the Deviation Form (Annexure-5).

6. Abbreviations

• SOP: Standard Operating Procedure
• CFU: Colony Forming Units
• QC: Quality Control
• TSA: Tryptic Soy Agar
• SDA: Sabouraud Dextrose Agar
• IPQA: In-Process Quality Assurance

7. Documents

1. Environmental Monitoring Log – Annexure-1
2. Surface Monitoring Log – Annexure-2
3. Personnel Monitoring Record – Annexure-3
4. Microbial Count Log – Annexure-4
5. Deviation Form – Annexure-5

8. References

• EU GMP Annex 1: Manufacture of Sterile Medicinal Products
• WHO TRS 961 Annex 6 – GMP for sterile pharmaceutical products
• USP <1116> – Microbiological Evaluation of Cleanrooms

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation	Microbiology Executive	QA Executive	Head QA
Department	Quality Control	Quality Assurance	Quality Assurance

11. Annexures

Annexure-1: Environmental Monitoring Log

Date	Location	Method	CFU Count	Analyst
24/06/2025	Filling Hood	Active Air Sampling	1	Rajesh Kumar

Annexure-2: Surface Monitoring Log

Date	Surface Area	Location	CFU	Analyst
24/06/2025	Gloved Hand	Filling Line A	0	Sunita Reddy

Annexure-3: Personnel Monitoring Record

Date	Employee Name	Location	Glove CFU	Chest CFU
24/06/2025	Vikram Singh	Grade A – Filling Line B	0	0

Annexure-4: Microbial Count Log

Sample ID	Date	Location	Result	Limit	Status
SP-AIR-2025-015	24/06/2025	Grade A Filling	0 CFU	Action Limit = 1 CFU	Compliant

Annexure-5: Deviation Form

Date	Area	Deviation	Immediate Action	Approved By
23/06/2025	Filling Zone B	2 CFU on Glove	Line Stopped, Re-gowning	QA Manager

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
01/01/2022	1.0	Initial Release	New SOP	QA Head
24/06/2025	2.0	Added updated alert limits and risk actions	Annual Review	QA Head