Steriles

Question 1

Does Annex 1 apply to non-steriles?

Answer 1

It can. If you claim to be following bits of Annex 1 for non-steriles then you need to document which principles have been applied and that compliance with those principles should be demonstrated

Question 2

What is a CCS and where should it apply?

Answer 2

CCS = contamination control strategy

Each facility must have a Contamination Control Strategy (CCS). This is to apply across the facility and includes pretty much everything (Annex 1 includes a list). It’s a living document that gets updated. Although control is individual items, the CCS hopes to look holistically at the full control system.

Question 3

What are prohibited in grades A and B?

Answer 3

Sinks and drains. Sinks are also prohibited in Grade B personnel change area.

Question 4

What does Annex 1 say about not using RABS or Isolators?

Answer 4

Alternative approaches should follow QRM and should be justified in CCS

Question 5

What design elements needed for steriles facility?

Answer 5

Transfer of materials into and out of an aseptic processing area should be single direction, ideally with a sterilising step incorporated (eg through the wall autoclaves and pyrogen tunnels.).

Airlocks should be separate for people and product. The at rest conditions in an airlock should be those of the clean area It is connected to. doors must be interlocked for grades A and B and considered for the other grades.

Positive pressure between zones (10 pascals) and flow visualised on a regular basis and videos maintained. Differential pressure monitors should be maintained and acted on when pressure goes out.

Viewing galleries should be provided.

Question 6

What do you need to clean RABS or isolators?

Answer 6

Cleaning of both RABS and Isolator should use a sporicidal agent and the post disinfection to batch start hold times should be validated (to ensure that the agent is gone).

Question 7

What are some of the tests needed for RABS and isolators?

Answer 7

Airflow studies will need to show the lack of ingress during events such as door opening. Glove leak tests need to be performed at the start and end of every batch. Need to do a visual inspection after an intervention and may need to do more, based on risk.

Note: article suggests all might not be lost if detect pin-hole. CCS should be able to help risk assessment – the main problem is when insert hands as pushes air into grade A. If operator does this directly over extract then could be low risk.

Question 8

What are the air grade requirements for RABS

Answer 8

Grade A inside / Grade B outside

Question 9

What are the air grade requirements for isolators

Answer 9

Grade D unless especially risky (refer to CCS)

Question 10

What are the air sampling requirements?

Answer 10

For classification, the number of sampling locations is as per ISO standard. However, for aseptic the grade A and grade B areas should consider additional sampling locations (etc stopper bowls / point of fill) in a documented risk assessment.

Question 11

What are the minimum requirements for requalification of clean room grade A

Answer 11

• Viable and non-viable particulates
• Integrity test of terminal HEPA
• Air volume
• dP between rooms
• Air velocity test

Question 12

What are the minimum requirements for requalification of clean room grades B, C and D?

Answer 12

Viable and non-viable particulates
Integrity test of terminal HEPA
Air volume
dP between rooms

Air velocity test as defined by risk assessment in CCS. Must do it for B background to RABS

Question 13

What are the requirements for disinfection?

Answer 13

Disinfection should be done according to a written programme and use two disinfectants, one sporicidal.

Monitoring should be performed periodically to ensure the programme is effective and detect changes to the background fauna.

Disinfection should be validated and disinfectants in grade A and B should be sterile.

Cleaning programme should remove disinfectant residue

Question 14

What are the requirements for particle counters?

Answer 14

Tubing <1m and bend radius >15 cm

Question 15

What considerations are there for maintenance in sterile facility?

Answer 15

If maintenance is done in clean area then should limit the number of people and define the activity to minimise contamination.

where unplanned activity / intervention is carried out then need to have an impact assessment to the rest of the product wrt sterility.

Foreseeable interventions should be validated.

Question 16

What are the requirements for inline filters?

Answer 16

Use 0.22µm filter for aseptic processes.

Filter to be integrity tested (PUPSIT) and results included in batch certification process.

Any pipework after the filter has to be sterilised.

Question 17

What are the requirements for personnel within the production area?

Answer 17

The number of people in manufacturing area should be minimised. The maximum number needs to be validated and confirmed during process simulation (broth trial).

People going into cleanrooms need gowning qualification and assessment and specific sterile training (micro / particle generation / hygiene / danger of getting it wrong). Gowning to be reassessed at least yearly. Each operator to do at least one process sim per year.

Duration of use of gowns in A or B to be validated.

Question 18

What do you do if the bulk can’t be sterilised

Answer 18

For bulk products that can’t be filtered then all inputs to be sterilised

Question 19

What hold times are required in a sterile facility?

Answer 19

There needs to be hold times for everything (cleaning to sterilisation, sterilisation to use, clean hold time of RABS etc, how long it takes to manufacture, how long parts are in the clean area before processing.)

Question 20

What inspection is required of parenteral products?

Answer 20

All filled containers of parenteral products should be inspected individually for contamination or defects. Defects to be classified and trended. OOT should be investigated. Need to have a defect library.

Automatic inspection systems have got to be shown to be as good or better than the human (who needs to be qualified and requalified annually)

Question 21

What should happen with open containers?

Answer 21

Open containers (partially lidded / stoppered vials / prefilled syringes) to be maintained in grade A with grade B background and separated from people using LAF carts.

Question 22

What are the requirements for steriles container closures

Answer 22

Containers should be closed by validated methods.

Fusion closed containers (ampules and plastic blow fill seal) to be 100% integrity checked.

Closures need validating which needs to consider worst case temperature and pressures

Question 23

What does Annex 1 say about sterilisation validation?

Answer 23

Sterilisation processes to be validated and efficacy tested with temp sensors and where appropriate BI. Validated loading patters need to be established for all sterilisation processes and should be revalidated.

Consider stress testing with max and min loads. At least annually for heat using the worst-case load pattern. Other load patterns should be validated at a frequency justified in the CCS.

Question 24

What is the minimum QC test for BI’s

Answer 24

If using BIs then need to confirm species of each batch. Can use C of C for other data (D-value etc)

Question 25

What happens to steriliser records?

Answer 25

steriliser records should form part of batch certification

Question 26

What are the general requirements for heat sterilisation?

Answer 26

Record each cycle

Probes to be in the worst places identified at Validation. Ideally checked against a second independent probe at the same position.

Beware cooling - need to maintain sterility. Any cooling liquid that touches the product needs to be sterile.

Question 27

What does Annex 1 require of moist heat sterilisation?

Answer 27

Typically steam but can use superheated water. Superheated water usually used for flexible containers where the pressure difference would damage the bag.

The items to be sterilised, other than products in sealed containers, should be dry, packaged in a protective barrier system which allows removal of air and penetration of steam and prevents recontamination after sterilisation.

For porous load cycles, each item should be dry on completion of the cycle and inspected post sterilisation for damage or moisture. If any is found then remove it and raise a deviation.

Weekly leak tests needed for vacuum cycle sterilisers. Daily test for air removal unless have an air detector system

Question 28

What does Annex 1 require of dry heat sterilisation?

Answer 28

Tunnels designed to maintain integrity of grade A.

Load pattern to be validated and probes in the worst-case area. Airflow patterns to be visualised and correlated to temperature studies. All air to be HEPA’d.

Critical parameters = belt speed / time, temperature, heat penetration, heat distribution and airflows.

Need to do 3 log reduction in endotoxins. For validation should use BI’s for endotoxins.

For ovens, all air entering the oven should pass through a sterilising filter. Parameters for Validation include temperature, time, pressure (as overpressured), air speed, air quality, heat distribution.

Question 29

What does Annex 1 say about EtO sterilisation?

Answer 29

Last resort due to toxicity. Need to worry about RH. Each cycle needs BIs. Need to pressure match before and allow time for EtO to be removed at the end of a cycle.

Validation parameters: EtO concentration, EtO gas pressure, EtO amount used, RH, Temp, Time

Question 30

What are the requirements around filter sterilisation?

Answer 30

The filter selected needs to be as described in the marketing authorisation. Consider second filter immediately before filling as part of the CCS

PUPSIT is mandated. If not possible then other approaches can be taken but need to demonstrate mitigates the risk. Filter should be able to be integrity tested in place. If have two filters then only need to test primary filter unless there’s a failure.

Parameters to be validated and monitored: wetting agent for filter integrity test should be as per manufacturers recommendation. If different to the original solution then it needs to not affect product quality.

Process conditions to be checked are: fluid pre filtering hold time, filter conditioning (prime with fluid), max filtering / contact time, max pressure, flow rate, max volume, temperature, time to filter known volume and pressure drop across filter.

Results of the above are to form part of the batch record.

Bioburden samples should be taken from the bulk and immediately before the filter.

Liquid filters to last 1 batch or 1 day whichever is sooner. Can extend if suitable validated.

Question 31

What are the requirements around sterile Form-Fill-Seal?

Answer 31

Grade A to be maintained in the critical zone and installed in Grade C environment (as long as Grade A/B garments are used)

Smoke studies and environmental monitoring needs to consider the moving parts and heat

For terminally sterilised Blow/Fill/Seal equipment should be installed in at least grade D. Filling conditions to be as per terminally sterilised stuff (i.e. C unless additional risk)

Question 32

What are the requirements around Lyophilisation?

Answer 32

lyophiliser to be sterilised and have a hold time. To be challenged during aseptic process simulations.

the frequency of vacuum / leak testing is to be documented and performed before every cycle. Integrity testing of the filter that maintains sterile conditions is to be part of the batch record.

Lyophiliser considerations:
* loading pattern documented
* transfer to be done under Grade A with as little operator contact as possible (LAF carts, HEPA tunnels)
* Airflow under all conditions to be maintained (inc transport devices and venting)
* unsealed containers (partially stoppered vials) to be maintained under Grade A (barriers expected). This includes where unsealed containers are removed from the chamber
* tools used during loading and unloading to be sterilised via a validated process.

Question 33

What are the requirements around sterile Closed Systems?

Answer 33

Sterility of all product contact surfaces to be maintained. Any post sterilising connections made must be aseptic connections.

May need integrity tests post assembly as per CCS

Question 34

What are the requirements around sterile Single Use systems

Answer 34

sterilisation process should be validated and shown not to affect the system. Verification of sterility should be part of supplier qualification and on receipt and use of each system.

Extractables and leachables should be assessed along with any interaction with the material.

Question 35

When should routine monitoring be performed and what should the limits be?

Answer 35

Routine monitoring to be done during all critical stages of manufacture, including set up. Needs to be done at the critical locations. Sampling methods should not pose a risk to the product.

Action and alert limits based on qualification / trend data and subject to review. Limits to be set such that a deterioration in conditions is detected early.

Only total particles for Grade A

Question 36

How do you determine the location of the EM requirements?

Answer 36

locations, frequency of monitoring, methods and incubation conditions should be defined by risk assessment.

Consider historical performance and background flora

Question 37

What are the four elements that an EM programme generally consists of?

Answer 37

This programme is typically comprised of the following elements:

i. Environmental monitoring – total particle.

ii. Environmental and personnel monitoring – viable particle.

iii. Temperature, relative humidity and other specific characteristics.

iv. Aseptic Process Simulation (aseptically manufactured product only).

Question 38

How often should the Grade A be EM’d?

Answer 38

Grade A is to be continuously monitored at ≥1 ft3/min. Need to check against limit above often to flag problems early. B wants to be the same but lower frequency.

Environmental and personnel monitoring - non-viable particles:

Need to monitor using settle plates, volumetric air sampling, glove, gown and surface sampling frequently but no impact the airflow patterns. Need to do personnel at periodic intervals during the process, especially after critical interventions and on each exit from the grade B cleanroom.

For grade A need to do continuous monitoring via air monitoring or settle plates and possible for grade B based on risk. Gloves and any other bit that may impact sterility should be monitored after critical operations as well as the exit monitoring. If it’s a manual process then it is one big intervention so frequency of monitoring needs justification based on risk (described in CCS).

people need to be monitored to assess aseptic behaviour.

Also need to monitor when not manufacturing.

Question 39

How are settle plates used?

Answer 39

Settle plates should be exposed in grade A and B areas for the duration of operations (including equipment set-up) and changed as required after a maximum of 4 hours (exposure time should be based on validation including recovery studies and it should not have any negative effect on the suitability of the media used).

• For grade C and D areas, exposure time (with a maximum of 4 hours) and frequency should be based on QRM.
• Individual settle plates may be exposed for less than 4 hours.

Question 40

Where do contact plate limits apply?

Answer 40

Contact plate limits apply to equipment, room and gown surfaces within the grade A and grade B areas. Routine gown monitoring is not normally required for grade C and D areas, depending on their function.

Question 41

When do you need to confirm the species of microorganisms?

Answer 41

For Grade A and B need to confirm species of microorganisms. Should also consider doing this for C and D but need to document your approach.

Question 42

How do you do a process simulation?

Answer 42

Need to use a media that grows the organisms. If your processing stages impact the viability (e.g. semi-solids, powders, heated or freeze dried product) then can use alternative procedure if justified.

Should assess the routine process as much as possible. Ideally, do the whole process rather than lots of individual assessments. If an inert gas is used then this should be replaced by air.

The process simulation should include known manipulations and interventions, including known corrective interventions that always happen.

Process simulation should be worst case for everything (times, people (inc longest time in suite), load, volume, fatigue, shift handover when equipment idle etc). Any gasses should be replaced by air unless anaerobic is being tested.

Question 43

What’s required for a media fill validation and when do you do them?

Answer 43

Three satisfactory consecutive media fills for validation that includes all shifts.

Need to do it after any significant modification. Or after a media fill failure.

Where manual operation (e.g. aseptic compounding or filling) occurs, each type of container, container closure and equipment train should be initially validated with each operator participating in at least 3 consecutive successful APS

Question 44

When do you do routine process sims and what’s involved?

Answer 44

Periodic process sims are done (usually twice per year) for each process, filling line and shift.

Each operator to participate in at least one successful process simulation per year. Also consider doing them before shutdowns.

Question 45

How big should your process sim be and what do you do with the containers

Answer 45

The number of filled units should be sufficient to effectively simulate everything it needs to. typically, at least 5000 to 10000 units. If smaller batch size than this then the process sim should be at least the same size as the production batch.

Filled containers should be swirled and tipped over to cover all sides of the vessel before being set down to incubate. The containers need to be clear so that can see any growth. Can substitute to clear for the test if necessary.

For active substances, batch size should be large enough to represent routine operation, simulate intervention operation at the worst case and cover all surfaces that may come into contact with the sterile product.

Question 46

What are the maximum action limits for viable particulates

Answer 46

Question 47

What are the airborne non-viable routine monitoring particulate limits

Answer 47

Question 48

What’s done in Grade A

Answer 48

Question 49

What’s done in Grade B

Answer 49

Question 50

What’s done in Grade C

Answer 50

Question 51

What’s done in Grade D

Answer 51

Question 52

What are the main differences between old and new Annex 1?

Answer 52

• Contamination control strategy
• People – more emphasis on behaviour and attitudes, gowning, laundering, qualifications, training.
• Cleanrooms – qualifying unidirectional airflow in grade A, and grade B to C airflow studies are required. Recovery rates of the room are required. Capture and compare at rest v in use.
• Improved section on transfer controls – protect grade A. Transfer should always be one way. Use validated sterilization or depyrogenation / disinfection. Sterile filtration of gases or liquids.
  o Prefer heat for sterilization – moist preferred. Secondary would be irradiation, filtration. If no other choice, ethylene oxide. SAY NO to UV-C, H2O2.
  o Filter as close to the vial as possible. Must control time, pressure, temperature, flow rate
• PUPSIT unless small volume and risk assessed.
• Media fill – detailed increased:
  o Acceptance criterion now zero growth
  o 3 consecutive repeats.
• Cleaning and disinfection increased – emphasis on cleaning (removal of residue), disinfection, rotation, use of sporicides periodically.
• EM needs to be dynamic, adaptive and dependent on the situation, e.g. out of shutdowns
• VHP – H2O2 deemed too fragile for a sterilizing agent. Non-seeking gas, sits on the surface.

Question 53

What is the ISO standard for cleanrooms?

What does it include

Answer 53

ISO 14644

Classification of cleanrooms by number of particles.
how to test
how to design

Question 54

You are manufacturing a sterile product. How would you go about generating an Environmental Monitoring plan?

Answer 54

Risk assessment of the production area that requires an EM plan. Include people flow, product flow, waste flow. Perform baseline monitoring of the area at rest.
Particle and microbial

MAP area at rest and take samples
MAP area in operation with a risk based decision on where to monitor
Choose routine samples

Question 55

What types of samples would you take in specific rooms e.g. change room, sterile room?

Answer 55

Particles, air, settle, gown