This is a ready-to-use qualification protocol for a cleanroom, covering the physical tests that earn the room its grade and demonstrate it holds cleanliness in the state the product actually experiences. Replace every <<FILL: ...>> placeholder, set your document numbers, and route it through document control. Set every numeric acceptance value from the current standard and your design, not from this template. A worked filled specimen follows. This content is educational and general; adapt it to your own quality system.
Approval page
| Role | Name | Signature | Date |
|---|---|---|---|
| Author (C&Q engineer) | <<FILL>> | ||
| Reviewer (HVAC/facilities SME) | <<FILL>> | ||
| Reviewer (Microbiology/EM) | <<FILL>> | ||
| Approver (QA) | <<FILL>> |
| Field | Entry |
|---|---|
| Protocol number | <<FILL>> |
| Room / area ID | <<FILL>> |
| Target grade / ISO class | <<FILL: e.g. Grade B, ISO 5 at rest / ISO 7 in operation>> |
1. Objective
To demonstrate that <<FILL: ROOM ID>> meets its classified air cleanliness and supporting airflow, filtration, and pressure requirements in the defined occupancy states, so it is qualified for its intended use in <<FILL: process>>.
2. Scope
This protocol covers non-viable particle classification (at rest and in operation), HEPA installed integrity, airflow velocity and/or air change rate, pressure cascade, recovery (for turbulent rooms), and airflow visualization. The viable environmental monitoring qualification is covered under <<FILL: EM protocol reference>>. It follows approved IQ (<<FILL>>).
3. System description
<<FILL: room use, grade, adjacent grades, HVAC arrangement, unidirectional or turbulent, HEPA locations, number and layout of monitoring points, barrier system if any>>
4. Prerequisites
| # | Prerequisite | Verified (Y/N) |
|---|---|---|
| 1 | IQ complete and approved; HEPA installed, gauges installed and calibrated | |
| 2 | HVAC balanced and stable | |
| 3 | Test instruments (particle counter, photometer/aerosol generator, anemometer, manometer) calibrated and traceable | |
| 4 | Classification diagram with sampling locations approved | |
| 5 | In-operation activity script defined (who does what) |
5. Roles
| Role | Responsibility |
|---|---|
| C&Q engineer | Executes or oversees the physical tests, compiles the report |
| HVAC/facilities SME | Owns the air system, balancing, filters, gauges |
| Aseptic operations | Provides representative operators and interventions for in-operation tests |
| QA | Owns acceptance criteria, reviews raw data, makes the qualified-state decision |
| Specialist test vendor (if used) | Performs certified tests on traceable instruments; report reviewed by QA, not accepted blind |
6. Tests, methods, and acceptance criteria
| Test | Method summary | Acceptance criterion (set your value) |
|---|---|---|
| 6.1 Particle classification, at rest | ISO 14644-1 count at approved locations, work height, isokinetic; sample volume per the standard | Each location at or below the class limit for each considered size |
| 6.2 Particle classification, in operation | As 6.1 during the scripted representative process activity | Each location at or below the in-operation limit for the grade |
| 6.3 HEPA installed integrity | Upstream aerosol challenge, downstream photometer scan of face and frame seal | No scan point above 0.01% penetration of the upstream challenge |
| 6.4 Airflow velocity (unidirectional) | Grid of anemometer readings at working plane | Uniform within the justified range (Annex 1 references 0.36-0.54 m/s at working position) |
| 6.5 Air change rate (turbulent) | Supply volume by hood/traverse divided by room volume | Meets the design ACR that supports cleanliness and recovery |
| 6.6 Pressure cascade | Differential across each adjacent boundary, doors closed | Each differential meets setpoint and direction; low-differential alarm functions |
| 6.7 Recovery (turbulent rooms) | 100:1 decay after a particle challenge | Recovery within the justified target time |
| 6.8 Airflow visualization | Neutral-buoyancy smoke, recorded, at rest and dynamic with interventions | Unidirectional sweep away from product; no in-draw over open product |
7. Test records
Attach a data sheet per test. Summarize results here.
| Test | Result summary | P/F | Data sheet ref |
|---|---|---|---|
| 6.1 Classification at rest | <<FILL>> | <<FILL>> | |
| 6.2 Classification in operation | <<FILL>> | <<FILL>> | |
| 6.3 HEPA integrity | <<FILL>> | <<FILL>> | |
| 6.4 Airflow velocity | <<FILL>> | <<FILL>> | |
| 6.5 Air change rate | <<FILL>> | <<FILL>> | |
| 6.6 Pressure cascade | <<FILL>> | <<FILL>> | |
| 6.7 Recovery | <<FILL>> | <<FILL>> | |
| 6.8 Airflow visualization | <<FILL>> | <<FILL>> |
8. Deviation handling
Any result not meeting its criterion is recorded as a deviation, assessed for impact on the qualified state, and resolved before the report is approved. Reference <<FILL: deviation SOP>>.
9. Summary and conclusion
| Field | Entry |
|---|---|
| Tests passed | <<FILL>> |
| Deviations raised / closed | <<FILL>> |
| Qualified recovery time recorded | <<FILL>> |
| Conclusion | <<FILL: room qualified to Grade/ISO ... in the stated states / not>> |
| Role | Name | Signature | Date |
|---|---|---|---|
| Executor | <<FILL>> | ||
| QA approval | <<FILL>> |
References
EU GMP Annex 1, Manufacture of Sterile Medicinal Products (2022 revision). ISO 14644-1:2015 (classification) and ISO 14644-3:2019 (test methods). ISO 14644-2 (monitoring) and ISO 14644-7 (separative enclosures, where a barrier is used). FDA Guidance for Industry, Sterile Drug Products Produced by Aseptic Processing (2004).
Describe these standards by number and title; do not paste their clauses. Confirm the current version before issue.
Filled specimen
Classification at rest, one room, to show the evidence level.
| Location | Particles/m3 at 0.5 um | Limit | P/F |
|---|---|---|---|
| B-01 | 1,210 | 3,520 | Pass |
| B-02 | 980 | 3,520 | Pass |
| B-03 | 1,540 | 3,520 | Pass |
| B-04 | 2,030 | 3,520 | Pass |
| B-05 | 760 | 3,520 | Pass |
| B-06 | 1,890 | 3,520 | Pass |
Six locations from the Annex A lookup for the floor area, sampled at work height, counter serial and calibration due date recorded, 1 m3 per point. Every location below the ISO 5 limit at 0.5 um; the room passes at rest. The in-operation run then repeats these locations during the scripted filling activity, and the smoke study films the same interventions, so classification, recovery, and visualization tell one coherent story.
Common inspection findings this protocol prevents
- Classifying only at rest, so the in-operation (worst-case) state is never demonstrated.
- In-operation counts taken with operators standing still, not doing representative work.
- HEPA integrity confused with filter efficiency certification, or the frame seal not scanned.
- Recovery time never established for a turbulent room, so at-rest and changeover times have no basis.
- A vendor report accepted without QA reviewing the raw data, calibration, or state represented.
How to adapt this protocol
- Set every numeric acceptance value from the current standard and your qualified design.
- Build the sampling location count and sample volume with the companion ISO 14644-1 worksheet.
- Script the in-operation interventions explicitly and reuse them in the smoke study.
- Tie the measured recovery time to a real operational rule (the post-exit settle period).
- Reference your IQ, EM, media fill, and requalification documents.