Performance Qualification is outlined below represents the quality testing performed in the OQ (i.e. viable) but expands the scope to include testing under dynamic conditions. The final and real challenge for the environment control system and HVAC system is represented by the process that must be executed within the areas it is serving. Upon determination of the new approved conditions, if needed, changes to the system are to be executed and revalidated before proceeding to the performance qualification.
Performance Qualification (PQ) protocol No PQP\VD\01\R00 shall be generated; the following qualification tests will be included in the PQ protocol and protocol must be prepared according to SOP-VD-012.
This section describes the various types of monitoring that would be performed to adequately qualify the operating environment for processing.
i. Performance Qualification must be performed on the facility in three different stages that are
i.1 “As-built” (No equipment, no personnel).
i.2 “At-rest” (equipment but no operations and no personnel) i.3 “Operational” (With personnel, equipment operations)
Consistent results must be obtained which should be within specified limits for 20consecutive working days for each of the three stages (as built, at rest and operation).
ii. Verification of Performance Qualification prerequisites
ii.1 The success of the system to control the level of viable and non-viable particulate levels as well as its ability to regulate temperature and relative humidity conditions depends not only on the system performance, but on outside factors such as personnel training, room sanitization, and system maintenance.
ii.2 Method for sanitization procedures, supporting utilities and personnel should be qualified/trained before the initiation of the PQ as well as procedures for sampling, operation and maintenance should be in place.
ii.3 By having these prerequisites completed before the HVAC system PQ and environmental monitoring program, the success for PQ is greatly extended and it is much easier to isolate attributable causes in the event of PQ failure.
ii.4 The prerequisites for the HVAC system PQ can be summarized below:
ii.4.1. HVAC: IQ and OQ complete.
ii.4.2. Utilities: IQ, OQ, PQ and including SOPs for compressed air and steam.
ii.4.3. Personnel: SOPs and training documentation for equipment operation, room sanitization gowning and environment monitoring. Training in pertinent SOP’s for HVAC & sterile area facility & all related operation and maintenance machines are completed and documented from all concern departments and persons
ii.4.4. Standard Operating System (SOP) must be development and approved before performance qualification (PQ) tests are executed for following:
a. Environmental Monitoring.
b. System Operation c. System Maintenance.
ii.5 All critical punch-list items from IQ and OQ must be cleared and resolved.
iii. Temperature – Humidity Control Test - Dynamic Condition
iii.1 To demonstrate the ability of the HVAC system to control temperature and humidity during operating conditions. This test must be executed while the process or operation are simulated or executed.
iii.2 The Temperature-humidity control test provides verification of temperature and humidity under dynamic conditions, as well as indicating that the system is capable of maintaining the design conditions. It also provides a good basis for determination of the general status of the system, for its malfunction can be used as diagnostic of the inadequate operation of the HVAC.
iii.3 Seasonal conditions of temperature and humidity may vary with the system design and the amount of external non-conditioned air supplied to the air-handling units serving the controlled environment. Sporadically, seasonal variations can be simulated during validation conditions.
iv. Differential Air Pressure and Direction Test – Dynamic Conditions
iv.1 In order to maintain air quality in critical and controlled areas, it is important that any airflow that occurs between adjacent areas which have different classification levels must be from the cleaner area to the less clean area (i.e., from a Class 5 area to a Class 7area). This is accomplished by maintaining the air pressure in the cleaner area at a slightly higher level than the air pressure in the less clean area.
iv.2 In s solid dosage facilities, airflow direction is designed to create conditions that contain product and minimize cross contamination. This is accomplished by maintaining the air pressure in the common area at a slightly higher level than the air pressure in the processing area.
iv.3 US guidelines require a differential pressure of 0.05” water column between rooms of different air cleanliness. EU guidelines require a differential pressure 10-15 Pascals.
iv.4 Differential Pressure/Airflow Directional Testing shall be conducted to verify the ability of the HVAC System to maintain plant pressurization positive between adjacent areas while in a normal operating condition (machinery in use and normal complement of operators present). This monitoring should be performed routinely and action levels should be established in the monitoring program per the requirements of ISO standard 14644-1 and 2.
iv.5 Differential pressure readings/measurements should be recorded using existing, (calibrated) installed system instrumentation. In the absence of such instrumentation, an independent calibrated dP gauge or inclined manometer with an appropriate range may be used. The recording of differential pressure measurements shall be conducted once a day for three (20) working days in order to demonstrate stability.
v. Air Cleanliness Test
v.1 Air Cleanliness test must be performed in order to determine that the complete as-built, operational facility meets the air cleanliness requirements specified in the user requirement specification.
v.2 Airborne concentrations are measured with white light, laser, or condensation nuclei particle counters.
v.3 Air cleanliness classification for controlled environments must be based on the concentration of particles of a specific size per unit of volume.
v.4 Sampling location and sample size must be set cautiously.
v.5 A sample should be taken at the fill point at a distance not exceeding 30cm (1ft3) from the point of exposure and a sample volume of 1ft3.
v.6 The table demonstrates the limit and range of air quality for every parameters that are performed in accordance with standard establishment
Parameters Limit/Range Reference TSI Instrument
Temperature 22 3C ASHRAE standard
55-1992
Q-Trak Plus IAQ-Calc TH-Calc
VelociCalc Plus
Relative humidity 30% to 65% ASHRAE Standard 55-1992
Ventilation (fresh air)
15 to 60 cfm/persons minimum depending on
Ventilation (CO2) About 700ppm over outdoor ambient
Ultrafine particles n.a n.a P-Trak
8 hr. TWA 1 hr TWA
vi. Airborne Bioburden Test
vi.1 The scope of this test is to determine if all the components of this system are capable of performing in accordance with the process functional requirements for airborne bioburden.
vi.2 The achieving of predetermined levels of microbial contamination in aseptic processing and supporting environments is a primary goal of the environmental control system.
vi.3 Suitable neutralizers should be added to avoid inadequate interaction with disinfectant agents employed in the environments.
vi.4 Organisms found in samples obtained from critical environments should be quantified and identified.
vi.5 Organisms found in other less critical environments should be quantified and identified also, in order to establish the prevalent flora in the environment.
vi.6 Selection of sampling sites is critical and special attention should be given to areas near personnel. As well as areas of transfer and connection to less critical environments.
vi.7 The sampling during validation must be extensive because the collection of microorganisms in such a clean environment is extremely difficult.
vi.8 Tests are performed to show that the air quality meets the specifications for particulates, temperature, humidity, microbial counts, lighting levels, etc. for the specification and classification of each room.
vii. Surface Bioburden Test
vii.1 The final evaluation of cleaning and disinfection must be performed in order to determine the bioburden content on the surface either in the facility or on particular equipment and device.
vii.2 The use of adequate cleaning and disinfection to achieve a level of quality environmental.
vii.3 As part of validation task, it is essential to determine that the effectiveness of this procedure is reflected at the rest of environment is being qualified.
vii.4 Evaluation of the surface bioburden test in pre-cleaning condition must be performed.
vii.5 Testing after the cleaning and disinfection in a repetitive approach, this must ensure to establish of these methods to perform such activity.
vii.6 Limits for surface bioburden test must be as follows:
vii.6.1. The limit for critical environment must be not more than 1CFU/12.9cm2 or 2 in2 (FDA Aseptic Processing guidelines).
vii.6.2. Similarly, other environments that are adjacent to critical environments: 5/12.9 cm2 or 2in2.
vii.6.3. For controlled environments: 20/12.9cm2 or 2in2.
viii. Non-Viable Particulate Testing (Critical and Controlled Rooms/Areas Only) – Dynamic Conditions
viii.1 Non-viable Particulate Testing shall be performed to verify the effectiveness of the Environmental Filters in minimizing and in effectively removing non-viable particulates, which may be present in critical and controlled areas.
viii.2 Testing shall be performed in accordance with the current U.S. FDA Guideline to Aseptic Processing and ISO Standard 14644-1
viii.3 Non-viable particulate samples shall be taken in each room or area in a normal operating condition (machinery in use and normal complement of operators present).
viii.4 Non-viable particulate samples shall be taken using an independent calibrated particle sampler and recording instrument, or may be taken using a qualified Particulate Counting System (PCS), if available.
viii.5 Each sample location within the room/area shall be sampled to determine the count of particulates. The number of sample locations shall be determined based on the floor area and the area classification.
viii.6 Limits:
Controlled Areas: The EU guidelines require these areas to be Class C.
Air in controlled areas is generally of acceptable particulate quality if it has a per-cubic-meter particle count of not more than 3,520,000 in a size range of 0.5 micron and larger (Class 8) when measured in the vicinity of the exposed articles during periods of activity. There is also a requirement to have less than 29,000 particles/m3 of 5.0 m and larger in the “in operation” state. To meet EU requirements both 0.5 and 5.0 m particle sizes must be counted.
Critical Areas: The EU guideline requires these areas to be Class A (corresponding to US Class 5 unidirectional flow), with the background of Class B. Air in the immediate proximity of exposed sterilized containers/closures and filling/closing operations is of acceptable particulate quality when it has a per-cubic-meter particle count of no more than 3,520 in a size range of 0.5 micron and larger (corresponds to US Class 5) when measured not more than one foot away from the work site, and upstream of the airflow, during filling/closing operations. It is accepted that sometimes conformity with particle counts may not be met at point of fill due to generation of particles or droplets from the product itself. Class B under dynamic conditions is of acceptable particulate quality when it has a per-cubic-meter particle count of no more than 352,000 of 0.5 micron and larger. The 5.0 m limits are 20 particles/m3 for Grade A and 2,900 particles/m3 for Grade B in operation.
ix. Viable Particulates Test (Critical and Controlled Rooms/Areas Only) – Dynamic Conditions
ix.1 The Viable Particulates Test shall be performed to monitor the viable particulates, which may be present in critical and controlled areas, and to determine the microbial quality of the air being supplied to each room or area.
ix.2 Viable particulate samples shall be taken in each room in a normal operating condition (machinery in use and normal complement of operators present).
ix.3 Sampling shall be conducted in accordance with the company’s procedures.
Sampling shall be performed for duration sufficient to sample each room during normal production.
ix.4 Each room shall be sampled a minimum of two (2) times per day, when the room is in production (operational condition).
ix.5 Testing should be performed over a time period of 20 consecutive days concurrent with the temperature, relative humidity, and differential pressure monitoring testing. In the event of no production in a room, a minimum of one (1) sample shall be taken for each location daily.
ix.6 Each room shall have no less than two (2) sample locations.
Controlled Areas: EU guidelines require a maximum average viable organism count of 100/M3 (2.8/Ft3).
Critical Areas: EU guidelines a maximum average viable organism count of <
1.0/m3 (0.03/Ft3), with a background maximum average viable organism count of /m3 (0.14/Ft3).
For testing to EU standards the average of the last 10 samples taken from a sampling site should be compared to the values given above.
Acceptance Criteria Test for Performance Qualification
Test Acceptance Criteria
Verification of
Performance Qualification prerequisites
All actions must be performed before starting execution of performance qualification activities.
Temperature – Humidity Control test
1. The specified temperature range must not be more than 22 3C.
2. The specified relative humidity range must not be more than 30-65% in aseptic processing areas. Unless otherwise specified by the process requirements.
Differential Air Pressure and Direction Test – Dynamic Conditions
1-∆P between any room and the main corridor must be within the tabulated defined range, which ensures that the ∆P between two adjacent rooms, which have same/different classification levels, is not less than 12.5pa. The supply and return air volumes should conform with the range specified. Pressure differential between rooms should be maintained as indicated in the specifications.
2-Absence of cross-contamination (measurement of pressure difference).
Air Cleanliness Test
1. Critical Environment: The particle concentration under dynamic conditions should not be more than 3.5 particles of 0.5µm and larger per cubic meter (100particles of 0.5µm and larger per cubic feet).
2. Other Environments: Typically a tenfold gradient must be used from critical to less critical environments (ex. 1,000 for environment adjacent to critical environment 10,000 for those adjacent to it.)
Airborne Bioburden Test
1. Critical Environments: Not more than 1CFU/m3 or 0.03 CFU/ft3.
2. Typical for other environments: Adjacent to critical environment 5/m3 or 0.15/ft3.
3. Controlled Environments: 87/m3 or 2.5/ft3.
Surface Bioburden Test
1. Critical Environments: Not more than 1 CFU/12.9 cm2 or 2in2.
2. Typical for other environments: adjacent to critical environments: 5/12.9 cm2 or 2in2.
3. Controlled Environments: 20/12.9cm2 or 2in2. Non-Viable Particulate
Testing (Critical and Controlled Rooms/Areas Only) – Dynamic Conditions
As defined in table, Airborne Particulate Classification (under point no. 6.3.5 in this report)
Viable Particulates Test (Critical and Controlled Rooms/Areas Only) – Dynamic Conditions
As defined in tablet: Recommended limits for microbiological monitoring of clean areas during operation (under point no. 6.7 in this report).
16. Planning and Cost: All planning and cost schedule is performed in Microsoft Project