Exam Questions

a) Z-value = Temperature change in °C required to alter D-value by factor of 10

• high z-value = means large increase in °C is required to achieve 1 log reduction of D-value = means that germs are more heat resistant to changes of temeprature

• low z-value = low increase in °C is required to achieve 1 log reduction in the D-value = germs are less heat resistant

  e.g. spores = z-value 10°C & vegetative cells = 5,5 °C

b) Relationship between the F value and the SAL value

F-value = thermal death time = holding time [mins] at a temperature T to reduce population N0 with specific D-value by desired number of poweres (log levels) to final value N

SAL = sterility assurance level = probability that germs may be alive

• Relationship: with sufficient F-value [holding time in mins] a sterilisation process can ensure that the SAL-value achieves the required standard

• high F-value = high holding time = low probability of items remaining non-sterile, low probability that germs stay alive = means a better (lower) SAL-value

• low F-Value = low holding time = high probability that sterilisation is not sufficient & germs stay alive = high SAL-value

(c) For a steam sterilization process at 134 °C, an F0 value of 50 min is achieved. How is the achieved F0 value of 50 min to be interpreted?

F0-value [mins] = 1 min means the lethality of a method that works for 1 minute at 121.1 °C (250 °F) and a z-value of 10 °C (18 °F) or an equivalent temperature-time combination

= makes different steam sterilization comparable -> important for pharmacy

50min (F0-value) at 134°C = process achieves the same microbial lethality as the sterilisation at 121,1°C for 50 mins

a) D-value = decimal reduction value [mins]

= time needed at a certain temperature to kill 90% of germs

= time required to reduce the number of viable organisms to 10 % of the original number

= time that is needed to reduce cfu number by factor 10 or by 1 log 10

Calculation:

Answer: D-value at 121 °C results 8 minutes.

b) Total time = F-value = time (holding time) at a temperature T in minutes to reduce the population (N0) with the specific D value by the desired number of powers -> often log10 levels (n)

n = from 100 germs to 10^-9 = 11 log levels

F (total time) = 11 * 8 mins = 88 mins

(a) How is the air removal carried out in the autoclave shown?

Steam generation within the chamber – single wall: Batch steam sterilizer

grid basket = Gitterkorb = air can circulated freely but can get unter the basket (enough space between basket & bottom needed for air elimination)

Function:

1) Product placed inside the autoclave

2) water reservoir is direct underneath the plate

3) steam generation with electrical heating (yellow)

4) Elimination of air = steam moves up & displaces the air to the top

 - air gets pressed out via air-removal valve out of the chamber

 - air must be eliminated otherwise steam cannot reach product; no condensation

(b) Name two disadvantages of the autoclave shown in (a) compared to an autoclave with an external steam generator.

picture a) Steam generation within the chamber – single wall: Batch steam sterilizer

disadvantages:

1) slow heat up because steam generation inside the chamber with (cold) product

2) slow cooling because water reservoir inside chamber, takes longer to cool system down (condensate stay inside)

(c) Draw the pressure curve of a steam sterilization process using fractionated vacuum for air elimination and a single vacuum phase for drying.

= combination of fractioned vacuum & single vacuum phase

A) Ventilation phase = generated steam replaces the air via air-separator

 = Risetime & air elimination:

B) Compensation phase = rest of the air gets eliminated

 = Pressure rises: valve get closed & steam accumulates

C) Sterilization time =

D) Cooling & vacuum phase = vacuum helps in drying

a) more than 500 containers (bottles)

2% of 10.000 = 200 -> its too much

• whihever is less: minimum 10 bottles for large volume parenterals should be tested

b) Procedure of sterility testing by by membrane filtration

Process: Sample filtration procedure

= should get done under cleanroom, laminar airflow work bench or in isolator

1) 2x empty bottles get connected with an injection needle a hose (Schlauch) through a peristaltic pump with the product bottle

2) Product extraction: with a peristaltic pump the product gets pumped out of the bottle -> product gets transported to the pre-sterilized bottles with a membrane filter

4) product is filtered & empty product bottle gets disconnected

5) a washing solution is placed via hose -> filters get washed to eliminate residuals (esp. of growth inhibiting product)

5) bottles get closed at the bottom

6) Steps for incubation: 2x different nutrient medium get introduced to the bottles to exclude (identify) different bacteria -> incubation for min. 14 days

(a) In which case may ionizing radiation sterilization be used for the terminal sterilization of pharmaceutical products?

= use of ionizing radiation possible for: pharmaceutical medical products

 dry powder products, non-aqueos liquids & semi-solids if the product is NOT temperature stable! -> e.g. ophthalmic ointments in tubes

• if product isn’t heat stable, use ionising radiation with high dose up to 25 kilo Grays

• Why is the used of ionising radiation limited? = can change the product characteristics due to the ionisation reaction

(b) Name one disadvantage of gamma irradiation compared to X-ray irradiation.

= Problem of handling/ disposing radio-active waste -> X-ray has no residues

= gamma-irradiation has lower oenetration capacity than X-ray

(a) According to GMP Annex 1, every company that manufactures sterile medicinal products requires a CCS. What does the abbreviation CCS stand for?

CCS = contamination control strategy (for Aseptic production of sterile drugs)

= a planned set of controls for microorganisms, endotoxin/pyrogen and particles, derived from current product and process understanding to assure process performance & product quality.

(b) One of the rules of aseptic working is not to interrupt the laminar air flow. GMP Annex 1 refers to this as the first air principle. Briefly explain why this rule must be followed whenever possible?

= is an important principle for aseptic working

• Laminar airflow shouldn't be interrupted because:

  1) Contamination control & Sterility maintenance: unidirectional flow of air sweeps away particles from working area -> when interrupted particles/ contaminants can settle on surface (risk of contamination) -> product quality & safety

  2) Important if protection of personal required (hazardous subtances) -> interruption can create a risk for personnel

  3) Laminar airflow is usually connected with controlled envirenmental conditions (°C/ humidity…) -> interruption can cause disturbances in conditions

(c) Which aseptic working rule should class B personnel follow regarding their gloves?

(a) Why is a safety cabinet considered less safe as an open RABS?

• at a safety cabinet operator is closer & in direct contact to working area, whereas in RABS interventions are made via gloves (barrier closed system) -> less safety of product & personnel in safety cabinet

• RABS enables higher level of sterility (due to enclosed barrier system, no room air & air moves from A to B), whereas safety cabinet is not enclosed (room air gets inside & filtered from B to A)

• Open RABS= vents in the barrier allow air to move from Grade A zone to Grade B area

(b) Which RABS type is most comparable to an isolator?

= closed (active) RABS = closed barrier between operator & critical zone, similar to isolator; glove portts are used

(c) When is overpressure used within isolators?

= for aseptic purposes, protection of product -> infeed/ fillig & part of loading area

(d) How can it be tested whether an inert gas atmosphere is maintained in an isolator?

= via Inertization test = test to verify that the establishment and maintenance of an inert atmosphere within the workspace is given by monitoring the concentration of oxygen (with oxygen meter) inside isolator

= also leak testing of Isolator can be performed e.g. oxygen sensor to avoid oxygen diffusion of isolator & maintenance of only inert gas inside

(a) Which medium is used for the final rinse in the washing process of primary packaging materials?

= Water for injection

(b) What is the key advantage of using single-use systems in aseptic manufacturing from a patient protection perspective?

= significant reduction of the risk of cross-contamination

= single-use is connected with purity/ sterility & therefore with safety for patient & quality of product

(c) What is shown in the figure below?

= a a sterile connector (hose connection with strile coupling)

(d) Why do syringes and cartridges receive an anti-friction coating on the inside?

= to minimize break loose & gliding forces (minimize friction)

-> without siliconisation on the indeside the stopper doesn’t move at all due to high friction or it moves in an uncontrolled way (no correct dosage of e.g. insulin possible)

(a) Why is weigh dosing the preferred method for aseptic filling for very high-priced products?

= enables high specification for dosing exactly to the defined amount

= to use exact ampunt of an high-prices product (economic aspect)

(b) What dosing system is shown in the picture?

= (sterile) Rotary piston pump

(18) Why must the monitoring results always be taken into account when releasing an aseptically filled product batch, even though a sterility test is carried out as part of the final product quality control? (1 point)

Monitoring / process control = Recording of process parameters temperature & time -> chemical & biological indicators

= to guarantee aseptic processing for the whole time of process

-> need to be proved that particle concentration is below limits with germ-free environment during & in the end of process

-> guarantees safety of process through recording of parameters & seeing if deviations may have negative impact of product quality (only good sterility testing not enough)

(a) An Ra value of 0.8 μm is specified for a surface. What does this mean?

(b) Name a method by which an Ra value of equal to or less than 0.8 μm can be achieved for a stainless steel surface.

• For stainless steel surfaces a Ra-value of ≤ 0.8 μm achieved by mechanical polishing or machining is recommended or via cold-rolling-process (Kaltwalzen)

(c) A material that comes into contact with a sterile product should be inert to the product. What else must the material be inert to? Specify one aspect.

= Inert (inaktiv) to the detergents and disinfectants

(a) What is meant by compensation time in (dry) heat sterilization processes?

Sterilization with dry heat: = At least 2 h at 160 °C -> Other time/temperature combinations possible -> increase of 10°C = doubles reaction process

= SAL: ≤ 10-6

Compensation time = additonal time required to ensure that the entire load (incl. difficult to reach areas) reaches the desired sterilization temperature.

= time that is needed to reach 160-180°C from the reference measuring point inside sterilizator & all points of product (even coldest spot)

= important to guarantee everywhere 121 °C

(b) The D value for heat-resistant endotoxins is around 50 °C. To achieve the required 3 log reduction of endotoxins the heat treatment is performed at 280 °C for 3 min. How long would the process duration be at 330 °C to achieve the same effect?

z-value = 50°C

T1 = 280°C

T2 = 330 °C

n = 3 log

t = 3min

DT2 = 2min * 10^ (280-330°C/ 50°C) = 0,3

F = 3 * 0,3 = 0,9 min = 54 sekunden

(f) How is the plastic tube produced in the BFS process?

(g) What is the minimum cleanroom class for filling terminally sterilized products?

= aseptic filling of terminally sterilised products is preferred to be done in class A & or at least in class C

(h) Bonus question: What is the name of the illustrated method for transferring tubs to class A, which requires double outer packaging of the tubs? Hint: the method is abbreviated NTT.

= Verarbeitung mit Doppelbeuteln, dem sogentannten No-Touch-Transfer (NTT)

= Dabei werden die Primärpackmittel steril angeliefert und über einen definierten Prozess, welcher kontinuierlich Keim- und Partikelüberwacht ist, in den aseptischen Bereich (Grade A) überführt.

= method designed to maintain the sterility of containers & components when transferring them into a critical cleanroom environment.

(d) Which steam sterilization process is suitable for the sterilization of bottled water for injections besides the hot water spray process?

= steam/ air mixture process

(e) The figure shows an autoclave tape. What is it used for?

2) Chemical indicators (for routine) / biological indicators (for validation)

-> chemical indicator tape = gets placed on product = shows colour change if desired conditions via sterilisation were reached -> indicator to controll success of sterilisation

(a) What type of filtration (in terms of separation limits) does sterile filtration belong to? Tick the correct box.

= is always the last option for sterilisation of pharmaceuticals

• Sterilization filtration (microfiltration) = separation of germs by filtration (sieving effect= Siebeffect)

• Membrane filter with an average pore size of < 0.22 μm (or smaller)

(b) Bonus question: Specify one common membrane material for sterile filters for the filtration of aqueous products.

(c) What is the unit of the bubble point?

Bubble Point [bar] = pressure at which a continuous discharge of air bubbles can be detected (3 to 5 bar) = depends on the filter properties

- big leak = lower pressure

- the higher pressure = the smaller the pore size

10^-5 bar = SI-unit is Pacal

(d) What is the acceptance criterion for the bacterial challenge test that is performed as one of the validation activities for a sterile filtration process?

= 100% retention of germs, no single reference germ should pass the filter!

(e) Sterile filters must be analyzed with regard to extractables, among other things. What are extractables?

b) microbiological tests of culture media (each batch):

1) Sterility testing = must be verified for each batch

-> incubation for 14 days & no visible microbial growth is allowed

2) Positive growth promotion test (with test microorganisms)

= proove that special test organisms can grow in the nutrient medium

-> Test principle: - very small number (100 CFU) of colonies is taken & placed in the nutrient medium (TBS or TM)

- different microorganisms (aerobic/ anaerobic/ fungi/ spores) get incubated to detect if they grow

In addition: biological test = with bioindicators (not culture medium)

c) Table Ph. Eur. 2.6.1

minimum volume per container - infusion solution (large volume parenteral) 250 ml

=> greater than 100ml = 10% of contents of the container (but not less than 20)

10% of 250ml = 25ml per container are used for sterility testing

d) How can it be ensured that the sealing point is also reliably cleaned and sterilised?

= due to CIP/SIP connection

= valve can get opened for cleaning and sterilisation at sealing point (orange front)

a) SAL = Sterility Assurance Level

= probability, that germs may be alive

= is expressed as the probability of a non-sterile item after sterilisation

= describes the probability of a unit being non-sterile after undergoing sterilization process

b) P = 0.99999. How many non-sterile containers would have to be expected for a batch size of one million containers?

SAL of 10^−6

= means a probability of not more than 1 viable germ in 1 ×10^6 sterilised items of final product

 = of 1 million product containers 1 container is contaminated after sterilization

Calculation:

SAL = 1-0,99999 = 1*10^-5

10^6/ 10^5 = 10

=> 10 non-sterile containers woulb be expected for a batch size of 1 mio. containers (with P = 0,99999)

a) under aseptic conditions ISO 5 (Class A) in isolator

= sterility testing isolator class A with surrounding area class D/C

b) Positive growth promotion test (with test microorganisms for each batch)

= prove that special test organisms can grow in the nutrient medium

-> Test principle: - very small number (100 CFU) of colonies is taken & placed in the nutrient medium (TBS or TM)

- different microorganisms (aerobic/ anaerobic/ fungi/ spores) get incubated to detect if they grow

c) acceptance criterion for sterility testing

= No visible growth (turbidity) after 14 days in the test sample

= a product is sterile if no microbes are detected in the sample after incubation

d) maximum number of containers of a parenteral

= 20 containers

(a) How do you call the release of terminally sterilized products without batch-related sterility testing, based solely on the measurement of physical process parameters?

(b) What must be considered in the PQ of a steam sterilization process with regard to loading - in addition to worst-case considerations?

= test load must contain a product that is routinely handled (loading patterns must be defined)

& the load must represent product families with the greatest difficulty for the sterilisation (worst-case consideration)

(c) Which third key specification of a bioindicator besides the z-value and D-value must be known for the validation of a steam sterilization process?

(d) What should not be included in the URS of a steam sterilizer? Mark with a cross.

URS = user requirements specifications & functional specification document

= a planning document that specifies what a system needs to do => part of DQ (Design Qualification)

• NOT included: = WFI-monitoring because use of steam

(21) The pipe transition shown (flow direction from left to right) has an unfavourable hygienic design. Draw a transition with an ideal hygienic design.

(c) How are glass ampoules sealed after the product has been filled?

= closure by fusion with flame or laser -> top of the ampoules froms by gravity

d) Which test must be carried out on all glass ampoules as a 100% check after sealing, regardless of whether the product is classified as a parenteral product or not?

= 100% integrity testing -> the integrity of the container closure system = include several tests (CCIT = container closure integrity testing)

• Validation of container closure integrity is important = prove with tests that sterility of product is maintained during storage

(e) You should plan a crimping process as part of a filling and closure line. Specify two aspects in addition to the supply of sterile caps that must be taken into account for the crimping process.

• Crimping of the cap should be performed as soon as possible after stopper insertion

• • while crimping vials a lot of metal particles get generated = non-viable particle

  -> therefore, important that crimping is separated from filling area (physical separation)

  -> measures to prevent particle contamination & good air extraction are required

a) Reference germs = test organisms

= are required for sterility testing to ensure the effectiveness of the test results

= for positive control: to verify that the sterilization process is effective in killing or inhibiting the growth of microbes

= to proove that test organisms cannot grow in the nutrient medium used for sterility testting or that the product doesn’t inhibit their growth

b) performance evaluation of the sterility test

= visual controll of turbidity after incubation of 2 weeks

c) Table 2.6.1

- more tha 500 containers

- 2% or 20 containers (10 containers for lang volume paranterals) whichever is less => 2% of 800 = 16 is less than 20

(a) A dose of 25 kGy is reached during irradiation. By how many log levels is a population of the yeast S. cerevisiae (D value: 0.5 kGy) reduced?

D = 0,5

25/0,5 = 50

= by 50 log levels reduction

(b) Name one advantage and one disadvantage of electron irradiation compared to gamma irradiation.

eletron irradiation

• Advantages:

  - No residues

  - More gentle than gamma irradiation

  - Continuous process, short irradiation time

  
  

• Disadvantages:

  - Limited penetration, but sterilization of products in final packaging possible

  - High dose rate

  
  

gamma irradiation

• Advantages:

  - High penetration capability for sterilization in final packaging

  - suitable for products with high density % large volume (e.g. whole pallets)

  - Rather low dose rate

• Disadvantages:

  - Problem of radio active waste

  - Not suitable for all materials (cross-linking / degradation reactions of polymers)

c) Why is UV-C radiation not suitable for the irradiation of e.g. medicinal products in sealed tubes?

= because absorbed on surface

= low penetration depth -> no reach of product

5) Why is "packaging material penetration" an important characteristic that a sterilization medium must fulfil?

= packaging material must be suitable for sterilization medium

• to ensure the efficacy & reliability of the sterilization process

• to ensure that every part of packed product get reached by sterilization medium (e.g. steam/ radiation)

• for qualified sterilization so that germs cannot survive

Typical heat killing curve of microorganisms (normal, non-logarithmic scaling of the axes) with a decimal reduction time of 1.5 min. (1.5 points)

• The F-Value has the unit … minutes

• The higher the F value, the … more extensive (better) is the killing of germs.

• The higher the z value, the … more resistant are the germs

• The lower the F value, the higher is the SAL value achieved (high probability of germs staying alive)

• With increasing temperature the D-value … decreases -> high °C results faster killing & a less D-value

• The F0 value is always connected with a temperature of … 121.1°C (for 1 min)

1. Higher z-value: Microorganisms are more resistant to temperature changes

2. Lower z-value: Microorganisms are more sensitive to temperature changes

   
   

1. Higher F-value: More extensive killing of germs.

2. Lower F-value: Less extensive killing of germs.

1. High F-value: More extensive killing of germs, resulting in a lower SAL value (higher assurance of sterility).

2. Low F-value: Less extensive killing of germs, resulting in a higher SAL value (lower assurance of sterility).

Increasing Temperature: The D-value decreases because higher temperatures increase the rate of microbial inactivation, leading to a shorter time required to achieve the same level of reduction in the microbial population.

a) What is the typical pore size of filters for sterile filtration?

< 0.22 micron (or smaller)

b) It is to be examined whether product is lost during sterile filtration, i.e. whether active substance is adsorbed in the filter. What is the procedure for this adsorption test?

Adsorption test = for sterile filterable products

= checking the amount of product loss; muss be 100% (no loss)

Procedure:

1) check change in concentration through sterile filtration

2) measure concentration of API before & after filtration

-> calculation of recovery = must be 100% no change in concentration

recovery = Wiederfindung: amount of product after & before sterilization = should be 100%

a) Why can sterility testing alone not guarantee the sterility of a batch?

• sterility means = absence of viable microorganisms

= because its not possible to test every container without damaging the product/ batch

= in reality only small sample (max. 20 sample size) get taken for testing -> there is always a statistical probability that the test might miss a contaminated unit if the contamination level is low

=> exponential killing (e^-x) of microorgamisms never reaches zero = never 100 % sterility possible -> D-value never zero

-> sterility has to be assured by the application of a suitably validated production process (geeignetes validiertes Produktionsverfahren)

b) product-specific suitability test

= to prove that the product doesn’t inhibit the growth of the 6 test microorganisms

=> product shouldn’t kill organisms that might me brought in a container during process (e.g. aseptic filling) to detect a contamination

2 Techniques of sterility testing = membrane filtration & direct inoculation

c) minimum volume per container - liquid vaccine 1.2ml

=> liquids 1-40ml = half the contents of each container but not less than 1mL -> half the contents 1.2/2 = 0.6ml

=> 1 ml per container must be testes

a) Why does pure steam have to be dry and saturated?

• Pure steam = pharmazeutischer Reinstdampf

  = steam with special purity, for direct exposure to group II sterilization goods

-> condensed pure steam has quality of water for injection (highest water quality)

• saturated steam (100% dry) = contains 100% of the latent heat available at that pressure

• Saturated water = has no latent heat & 0% dryness, contains low heat.

= has to be dry & satured for effective sterilization:

1) Optimal heat transfer = dry satureated steam has highes amount of energy and can transfer this energy more effectively (weat has lower energy transfer is & takes too long)

2) can condensate immediately -> because saturated team has boiling point of water ensuring a concitent temperature (better effectiveness)

3) with dry saturated steam germs are exposed to the max. thermal energy leading to more reliable sterilization (as with via wet steam, droplets)

(b) Why is there also a limit value for bacterial endotoxins in pure steam?

  = pure steam can get in direct contact with pharmaceutical products

(e. g. primary packaging, solutions for production of aseptic medicinal products, parts in contact with medicals)

-> limit value to ensure that product is not cntaminated -> to ensure patient safety, product integrity, quality & positive (desired) clinical effect of the drug

Describe the complete sequence of sterilization using ethylene oxide.

= generally, only acceptable if no other method of sterilisation is possible” -> use in food sector prohibited

= in pharma limits of dosage are defined due to toxic residues

-> is very explosive when in contact to O2

Process sequence (negative pressure process)

= runs under vacuum because of 2 purposes:

I) test of integrity of sterilization chamber

 II) to take out air because it forms explosive mixture with ETO

(a) What is the difference between a passive and active RABS?

• Active RABS = integral HEPA-filtered air supply (HEPA inside)

• Passive RABS = air supply by sealing mounted HEPA-filters (outside)

Open, active RABS = HEPA filter is inside, independent air supply unit

Open, passive RABS = air flows aout of the system, not for toxic products

(b) Which cleanroom class must be used as the background environment for a RABS?

• Surrounding room classification should be ISO 7 minimum in operation -> surrounding area of RABS = class B

Isolator surrounding = for open isolator class C & for closed isolator class D

(c) What are the characteristics of an isolator called "open aseptic"?

Product protection isolators = “Aseptic isolator”

II) Open isolator systems = allows continuous material flow, hermetic seal for decontamination, with unidirectional air flow

 = designed for continuous or semi-continuous ingress and/or egress of materials (Ein-/ Austritt) during operations through one or more openings -> mouse holes

 = Openings must be engineered (e.g. using continuous overpressure) that no external air/ contaminant get into the isolator

-> has tiny openings (“mouse holes”) with inside overpressure to ensure that no particles get inside

-> min. grade C surrounding (because of holes)

(d) How do you perform the leak test of an isolator glove (in addition to the visual inspection)?

2) Leak testing:

 2.1) Pressure decay test = gloves get blowed up with pressure

 2.2) airflow test at constant pressure = air gets inside of gloves & measurement of volume that is needed to keep pressure constant => if tight, no additional air needed

a) Name two approaches to prevent particles / microorganisms from entering the aseptic area from the glass washer area in addition to the high temperature

1) pressure cascade

2) Forced air circulation with HEPA filtered hot air (laminar airflow (LAF) = to remove particles & to control pressure conditions

b) What is the objective (Ziel) of the higher temperature in the pharmacopoeia?

= to ensure a more effective sterilization process -> Increased Sterilization Efficacy, high sterility assurance

= to enable depyrogenation which starts above 220 °C (for glass) to reduce heat resistant endotoxins

= to fulfill all Pharmacopoeial Standards for sterilization -> product & patient safety

a) Which prerequisite must be fulfilled for an aqueous medicinal product to be prepared aseptically at all?

1) sterile material & equipment

2) class A conditions (environment)

3) sterile transport

4) Isolator

5) sterile product

b) The condition from (a) is given and a drug is to be prepared aseptically. However, it cannot be sterile-filtered.

What is the aseptic production process in this case?

= product is not heat stable at all & cannot be filtered then: use pre-sterilised individual or mixed components & aseptic processing

= all product components need to be sterilized separately beforehand & bring/ fill everything in aseptic processing together (Aseptic formulation: class A)

Important: Terminal sterilization is considered as much more saver as the aseptic processing because final product is completely closed in the final packaging. It cannot get contaminated because of the validated container closure integrity

(a) Describe the principle of a transfer using a decontamination airlock into an isolator during operation.

Decontamination airlock (Luftschleuse)/ transfer hatch

= has 2 decontamination airlocks (small chambers) on each side of the isolator that are connected to working area

-> material can get inside & out of the isolator

-> double door system

(b) What transfer system is shown in the picture?

RTP = Rapid Transfer Port

= for transfer of materials, stoppers, caps into aseptic area while processing

How can the open product in cleanroom class A can be protected from contamination?

1) physical protection

- Walls/ Barriers

- Shell concept: surrounding class B

2) aerodynamic protection

- protection via laminar air flow

- pressure cascade between zones

3) Personnel (human factor) = try to avoid human interaction through using automatic system

4) Use materials (incl. API & excipients) with very low bioburden

5) Surfaces in production surrounding = compatible hygenic design -> quality/ cleaning/ roughness for desinfection

a) What is checked during the OQ of a steam sterilizer?

OQ = Operational Qualification = Checking basic functions of equipment within the working area including chamber integrity

-> checking if:

1) sterilization possible

2) are required °C possible to achieve

3) chamber integrity -> possible with vacuum

b) What is the general name of test strips with bacterial spores used to validate sterilization procedures?

= biological indicator (microbial tests)

c) What should not be included in the URS of a steam sterilizer? Mark with a cross.

DQ = Design Qualification = Determination of requirements for equipment

  URS = user requirements specifications & functional specification document

a) pure steam = has special purity & high quality so it can get in contact with pharmaceutical

-> if there is a lot of air or non-condensable gases inside sterilization system the steam cannot reach every area of product

=> results: bad sterilisation quality

- no condensation possible

- bad energy transfer -> needed °C cannot be reached for desired duration

b) Advantage of saturated steam = condensates quickly -> shorter process time

(super-heates steam = needs to cool down to condensate -> longer)

a) What D value at 121 °C results from the data?

b) After what total time is a SAL of 10^-12 achieved if 10000 germs are present at the beginning?

(a) Why is primary packaging material considered more critical then secondary packaging material?

= its the first layer of packaging that is in direct contact with pharmaceutical product (e.g. vials/ ampoules/ syringes/ stoppers/ caps)

-> shouldn’t cause recontamination or have negative impact on product -> must be sterile (depyrogenation) / inert/ abrasive proof etc.

-> Sterilization tunnel (continuous sterilization) = used for sterilization & depyrogenation of primary packaging materials (glass containers) -> not for plastic because high °C!

(b) Name one way in which two silicone tubes can be connected aseptically.

(c) Why are cartridges siliconized on the inside?

• without siliconization stopper doesn’t move at all due to high friction or stopper suddenly moves & stops -> syringe doesn’t work well, no correct dosage (insulin) -> dangerous for patient

break loose force = force that is needed to start movement of a stoper

gliding force = force that is needed to keep stopper moving

(d) At which point in the process does siliconization take place?

1) siliconisation of packaging (syringe/ cartridge) usually before sterilisation (after washing & drying) -> often applied backed on siliconization which has formation of silicon layer during (hot air) sterilization

2) siliconisation of syringe & cartridge stoppers = optionally after sterilization -> stoppers must move to achieve equal distribution of silicone

a) (Dual) peristaltic pump

= single-use silicone tube -> tube get sandwiched between rollers (Dialyse-pumpe)

= benefits: single use/ high precision/ quick product change possible

(b) Explain the principle of time-pressure dosing.

Time-pressure system = gravity & viscosity dependent system

- benefit: ultra-high output/ CIP-SIP compartible

- not for: viscous products & products that are sensitive to silicone tubes

Principle:

- pressure inside vessel must stay constant -> precise monitoring of pressure

 - pinched valves are opened & then after defined dosed product valves get closed

 - temperature very important (high impact on viscosity) -> influences flow through

(c) Why does Annex 1 specify that crimping must be separated from filling and stoppering?

Crimping = aluminium cap gets placed on the stopper

• while crimping vials a lot of metal particles get generated -> non-viable particle => therefore, important that crimping is separated from filling area (physical separation)

=> to prevent particle contamination & good air extraction are required

(d) Which test must be carried out on glass ampoules after fusion as a 100% check in addition to the visual inspection?

-> closure by fusion, e.g. glass or plastic ampoules should be subject to 100% integrity testing.

(e) What is the main advantage of using nested ready-to-fill syringes in tubs compared to using bulk syringes?

bulk syringes = Washing, drying, siliconisation, sterilization necessary before filling

nested ready-to-fill syringes = just opening & filling & closure (ready to use) -> no washing & sterilization etc.

(f) How do you call the process for manufacturing infusion or parenteral nutrition bags? = form- fill- seal production

(g) What is used to blow up the containers during the blow fill seal process?

Blow/ fill/ seal system = purpose-built machine in which, in one continuous operation, containers are formed from a thermoplastics granulate, filled & sealed, all in one production line

Blow mould closes, bottom is welded -> blowing with sterilized compressed air

a) Draw in a design improvement for the pictured container bottom.

(b) Name two criteria for the design of pipework that are not related to the properties of the material / surfaces.

• Self-drainable -> have special slope (are not straight)

• Fixed connections prefered to detachable ones

c) Sketch / draw the compression of a seal with an axial stop

d) Which one is better with regard to hygienic design / contamination risk? Please briefly justify your choice.

Sealing of longitudinal movements

a) dynamic sealing

b) better: hermetic sealing = because hermetically sealed & no germs can enter

(e) Why is a simple mechanical seal not suitable for sealing agitator shafts?

= is NOT 100% sealed = not 100% safe: contamination get through sealing point

-> cannot be used for aseptic purposes & hazardous media -> used for non-sterile/ non-hazardous product/ pumps -> microbial contamination can get through sliding point

a) What must be considered in addition to the routine process when planning a media fill?

Media fill = should imitate as closely as possible the routine aseptic process & include all critical manufacturing steps -> microbial monitoring

= should consider various interventions known to occur during normal production & worst-case situations -> risk analysis

b) In a media fill, 600 containers are filled with nutrient medium. How many of them are allowed to show germ growth after incubation?

= no growth (0 germs/ growth) -> otherwise something is not right in production

c) Which special aspects have to be considered for a media fill if a freeze-drying process is part of the procedure to be validated?

Lyophilizates -> partial drying process

- FDA recommends that unsealed containers be exposed to partial evacuation of the chamber that simulates the process.

- Vials should not be frozen, and all precautions should be taken so that microbial growth get supported (for evaluation)

(a) Media fill is also known as APS. What does APS stand for?

APS = aseptic process simulation

(b) How often must a media fill be performed as part of the introduction of a new product?

for new product = initial validation = 3x successful APS

& periodic revalidation 2x/year

(c) An SOP for the aseptic formulation and filling of a liquid product specifies that two excipients and WFI should be added to the active ingredient while stirring gently. How do you take these specifications into account for a media fill?

= instead of product is a nutrient medium used

= media fill hast to be performed according to defined aseptic process according to manufacturing SOP & media fill plan:

• Simulation of process still necessary: = Use Placebo Excipients (instead of the actual excipients) -> goog planning necessary that placebo do not affect microbial growth. -> proper documentaton/ monitoring etc

(d) How long does incubation take after a media fill and how are the incubated containers subsequently analyzed?

= incubation for 14 days

Evaluation = via visual inspection & turbidity

9) Describe the steps that are needed to biodecontaminate a cleanroom with hydrogen peroxide vapour (VHP process)?

(a) What is meant by mouseholes in barrier systems and for what purpose are they used?

b) When using RTP systems, how is it ensured that no contaminated air enters the barrier system (see illustration)? Specify two aspects

2) connection must be air-tight to avoid decontamination

3) opening of lid only possible when fully connected

4) higher pressure inside transfer pressure for product protection

(a) Which D value at 121 °C results from the diagram?

calculate: D-value = mins/ log-levels = 8mins/ 4 log levels = 2 mins

or read in diagram

b) What is the D value at a temperature of 131 °C?

What data are needed for validation with biological indicators?

1) bioburden N0

2) type of microorganisms -> resistance: D & z-value -> survival & kill-time

Why is sterile compressed air for counterpressure used and not steam?

 = because to generate steam higher °C is needed which raises the pressure: p = p1 + p2 -> container may burst

a) What does the SAL value describe?

SAL = sterility assurance level = probability, that germs may be alive

SAL of a sterilation process = degree of assurance with which the process makes a population of items sterile

= is expressed as the probability of a non-sterile item in that population

b) What is the probability of sterility achieved if the bioburden before sterilization is 20 cfu/ml in a 5 ml volume and sterilisation results in a 7 log reduction?

a) The following z value (temperature coefficient) is given in connection with a steam sterilization process: z = 6 °C. What is the D value at 121 °C if the D value at 109 °C is exactly 30 s?

c) Formula cannot be used because no z value given!

What role does the bioburden of the sterilization items play in the design of a sterilization process?

• the lower the bioburden that easier is it to guarantee sterility

• reduction of bioburden includes washing/ cleaning/ desinfection & sterilization in the end

• The level of bioburden affects the required lethality of the sterilization process. Higher bioburden = requires more strict sterilization parameters (e.g., higher temperatures, longer exposure times) to ensure good microbial killing

• Knowledge of bioburden helps to select the appropriate sterilization method (e.g., steam, dry heat, radiation) and optimized parameters such as temperature, pressure, and duration to achieve the desired Sterility Assurance Level (SAL).

a) SAL + Psterile = 1 = 100%

SAL = probability, that germs may be alive

Psterile = probability of germs that are death

b) 10^-5 = 1 container of 100.000 is non-sterile

1,500,000 containers * 10^-5 = 15 containers are non-sterile

c) acceptance criterion for the SAL value

= min. SAL of 10^-6 for pharmaceuticals

-> in reality pharma companies chieve SAL of at least 10^-12

a) For which pharmaceutical products can the hot water spray process be used?

Hot water spray = Counter pressure sterilization processes

= for aqueous liquids (infusion) in closed containers

= e.g. for bottles, vials made of glass/ plastics & single-multi bags

(b) How is the counterpressure built up in the hot water spray process to prevent the product containers from bursting?

 = via controlled application of compressed air in the chamber -> to balance the internal pressure inside the containers with the external pressure in the sterilization chamber

• As the temperature inside the chamber rises, the pressure also increases. To prevent the containers from expanding or bursting due to the increased internal pressure from the heating, compressed air is introduced into the chamber. => The introduction of compressed air increases the external pressure in the chamber, creating a counterpressure that balances the internal pressure of the containers.

(c) In which steam sterilization processes is the Bowie-Dick test used?

= for saturated steam sterilisation with vacuum processes

= proof if sufficient air removal achieved & steam penetration into product sufficient

a) What does this z value of 5.5 °C indicate?

z-value of 5,5 °C = Temperature change of 5,5 °C, which is required to alter the D value by a factor of 10 -> 1 log reduction

= measures the heat resistance of microorganisms

= low z-value means that germs are sensitive to °C (e.g. vegetative cells are not heat resistant)

b) How are the F0 value and the SAL value related?

F0-value = special F-value for 121 °C conditions

• the higher F-value (long holding time) = the lower SAL-value (low probablity that germs stay alive, sufficient sterilisation)

• the higher the F-value (short holding time) = the higher the SAL-value (high probablity that germs stay alive, no sufficent sterilisation)

a) in hot air tunnel:

1) Depyrogenation (while sterilization phase) = to reduce heat resistant endotoxins

2) seliconization = silicon layer is spread / made up to reduce friction (for glass or in syringes)

b) Give a common operating temperature for the sterilization zone.

= 280 - 350°C

c) pressure zones in hot air tunnel:

1) washing zone = +2 Pa (refer to class C pressure)

2) heating zone = +4 Pa (sterilization)

3) cooling zone = +6 Pa (class A conditions) = to make shure that no air is getting inside sterilization tunnel

(a) Explain the effect of air pockets within the sterilization load during steam sterilization.

Steam sterilization (= autoclaving) relies on the presence of saturated steam to transfer heat efficiently to all parts of the load.

-> when saturated steam is coming into an autoclave = it reaches a cold product surface & gets condensate because the steam cools quickly down & transfers all its saved energy to the product -> so product heats up

-> condensation = very fast & powerful energy transfer process

Effects of air pockets:

1) incomplete sterilization: - Air pockets act as barriers to the penetration of steam -> due to air pocket area may not reach the required temperature for effective sterilization -> creation of cold spots -> higher risk of surviving germs -> air prevents steam for reaching the product -> bad sterilization

2) longer sterilization time: - Steam provides higher efficient heat transfer than air -> areas with air pockets require longer exposure times to reach the required temperature -> extention of overall cycle time

b) How is the pressure in an autoclave related to the amount of noncondensable gases in the autoclave chamber?

limit < 40ml/kg non-condensable gases (NCGs)

3) Pressure increase: In a closed system the pressure rises up (because of high energy & steam needs more space) so the vaporization curve reaching 212°C and it becomes a saturated steam

= if pressure rises to high, then there is a lot if air left causing overpressure (p1+p2= Ptotal)

Effects of Noncondensable Gases on Sterilization

1. Interference with Steam Penetration: NCGs can form pockets or layers that disturb the distribution of steam, leading to uneven heating & insufficient sterilization.

2. Lower Effective Sterilization Temperature: The presence of NCGs can lower the effective sterilization temperature. For example, if the chamber pressure is read as 121.1 °C (1 atm of steam pressure), but part of this pressure is due to NCGs, the actual steam temperature will be lower than 121.1 °C. -> reduced SAL & uneffective strilization

(c) How is venting / air elimination performed in a steam sterilization process that uses the gravity or flow principle?

(a) Name two further requirements in addition to easy cleanability & smooth surface for materials in contact with the product with regard to their hygienic design.

Requirements for materials - hygenic design

1) Smooth, easy to clean

2) Corrosion resistant -> stainless steel

3) Abrasion-proof, mechanically stable

4) Stable to aging & not change over time -> rubber may break in peaces

5) Inert (inaktiv) to the product & detergents and disinfectants

b) What is designed in a hygienically unfavourable way here (see picture)? Name one point of criticism.

- Sharp corners (≤ 90°) must be avoided

- surfaces hall always slope to one side with a minimum angle of 3°

- not self-draining

1) visual inspection is tiring for eyes -> looking 5 secons & making regular pauses

2) Personnel should follow hygiene practices, to prevent contamination of the products during visual inspection

3) Adequate and good lighting conditions are important for effective visual inspection

4) personnel friendly working condition (temperature/ humidity) are important -> creation of particles through sweating/ low concentration

a) What is meant by compensation time in (dry) heat sterilization processes?

b) In addition to sterilization and depyrogenation, what third purpose is met by the treatment in the hot air sterilization tunnel?

= siliconization : silicon layer (anti-friction coating) is made up to minimize break loose & gliding forces

-> for cartridges & syringes = have stopper inside, it has to move while injection

-> silicone oil or emulsion is sprayed to the inner surface of syringe

(c) The z value for heat resistant endotoxins is about 46 °C. What does this mean when you compare the required process times at 280 °C and 326 °C?

t = 326-280°C/ 46°C = 1

=> 1* z-value (reduction of 10log)

= Process tima at 326°C will be reduced by factor of 10 compared to 280°C

d) Why is it important to cool glass containers slowly after hot air treatment?

= to prevent thermal stress and potential breakage

= to avoide class breakage (due to possible micro-cracks)

= to guarantee product safety (no contamination through glass crackes)

a) D-value = mins/ log levels

13 -1 = 12mins -> 12/8 (log-levels) = 1,5 D-value

b) D-value = 1.5 -> n = log-levels: from 10^8 to 10^-6 = 14 log levels

1.5 * 14 (log levels) = 21 minutes

c)

The F0- value is reached below 13 minutes at 125 °C.

The sterilization process at 125 °C achieves the required F0​ value much earlier than 13 minutes.

a) SAL + Psterile = 1 = 100%

 SAL = probability, that germs may be alive

Psterile = probability of germs that are death

1 - P = 0,00000002 SAL = 2*10^-7

b) Is the SAL value achieved sufficient for a pharmaceutical steam sterilization process or not? Please justify.

-> in pharmacy = aim to achieve SAL level of equal to or less 1*10^-6

= is achieved & even axceeded because SALof 2*10^-7 means that 2 containers of 10 Mio are unsterile (1 unsterile container of 5 Mio)

-> SAL of 1 * 10^-6 means 1 container of 1 Mio unsterile

a) SAL 1.8 * 10^-11

Redution: 13 log steps -> 11.8 * 10^(-11+13)

initial baterial count N0 = SAL + log steps = 1.8 * 10^2 = 180

b) Probability in percent:

SAL + Pst = 1

Pst = 1 - SAL = 1 - (3 * 10^-2) = 0.97 = 97%

(a) In which case may ionizing radiation sterilization be used for the terminal sterilization of pharmaceutical products?

= if it is NOT possible to sterilize pharmaceuticals by heat or steam (not heat stable product), then ionizing radiation sterilization may be applied

= if product cannot be sterilised by dry heat at 160°C for 120 min or by any other alternative temperature achieving SAL < 10^-6, the use ionizing radiation

-> for: pharmaceutical medical products; dry powder products, non-aqueos liquids & semi-solids if the product is NOT temperature stable!

 e.g. ophthalmic ointments in tubes

(b) According to which mode of action does UV radiation, which is not one of the ionizing types of radiation, kill microorganisms?

• Damages DNA (maximum at 265 nm) -> kiling of germs due to disruption of DNA replication & transcription

= DNA is absorbing UV-energy causing apoptosis of cells/ mutations

(c) Which of the ionizing radiation types does not belong to the electromagnetic radiation?

= alpha & beta rays

a) What was the initial bacterial count N0 before sterilization if this SAL valuewas achieved after a reduction of 8 log steps?

5* 10^-7+8 = 5*10^1 = 50

b) What is the probability of sterility in percent associated with the SAL value?

(a) What is the common mechanism of action of all types of ionizing radiation sterilization?

= induction or damage to the genetic material (DNA or RNA) of microorganisms (indirect or indirect way)

- Ionizing radiation has enough energy to ionize atoms/ molecules , leading to the formation of highly reactive ions / free radicals. These can then interact with and damage the genetic material of microorganisms, making them unable to reproduce & causing their destruction.

(b) For pallet sterilization of consumables, a radiation sterilization process with the highest penetration capacity should be selected. Material compatibility is given. Which type of radiation can be considered besides X-ray?

Gamma irradiation

= High penetration capability sterilization of products in final packaging

= suitable for products with high density & large volume (e.g. whole pallets)

(c) Name an area of application for decontamination with UV radiation.

= in water treatment -> used to disinfect and decontaminate water

a) Which two irradiation methods are thus applicable for terminal radiation sterilization (Annex 12)?

1) electron irradiation

2) gamma irradiation

b) Irradiation for sterile transfer

= E-beam tunnel -> transfer from class C to class A

(a) What is the mechanism of action of gamma irradiation?

= use of high-energy gamma rays to disrupt the DNA or RNA of microorganisms, making them incapable of reproduction

• mechanism of action:

  1) Generation of Ionizing Radiation: = cobalt-60 creates during radioactive decay high-energy gamma rays

  2) Penetration of Materials = Gamma rays have high energy and short wavelengths, allowing them to penetrate deeply into materials (sterilization)

  3) Ionization of Atoms & Molecules = As gamma rays pass through a material, causing ionization of atoms -> results formation of highly reactive ions & free radicals -> killing germs

(b) Give a reason why gamma irradiation is preferred over electron irradiation for this application.

electron irradiation = Limited penetration, but sterilization of products in final packaging possible -> may be not sufficient to penetrate whole pallet (bad sterilization)

Gamma irradiation = High penetration capability sterilization of products in final packaging -> higher assurance level (to penetrate whole pallet)

(c) Name one advantage and one disadvantage of gamma irradiation compared to sterilization with ethylene oxide.

Gamma Irradiation

• High penetration capability sterilization of products in final packaging & no residuces

• Problem of radio active waste

Advantages - ETO

1) Broad spectrum of activity

2) Alkylation of nucleic acids, proteins (requires H2O)

3) Excellent penetration capacity

Disadvantages - ETO

1) Carcinogenic

2) Extremely flammable, forms explosive mixtures with air

3) Leaves toxic residues -> not allowed in food sector

(d) Give a reason why ethylene oxide is used for that application but not hydrogen peroxide = sterilisation of entire pallets with consumables.

because: ETO has excellent penetration capacity & broad spetrum of activitity -> better for entire pallets with undefined consumables

hydrogen peroxide

= Very strong oxidizing agent

= Highly corrosive

= Penetration capacity limited

a) Why are hot air sterilization tunnels usually operated at temperatures of 300 - 350 °C?

1) to eliminate pyrogens (depyrogenation) -> above 220 °C

2) because its a (faster) process (sterilization cycle) resulting better killing & higher sterility assurance

b) Which zone of a hot air sterilization tunnel has the lowest ambient pressure?

= the drying/ preheating (infeed) zone (class c) = +2Pa

-> highest pressure in cooling zone (after sterilization to ensure sterility)

c) Why is it important that glassware cools slowly after hot air sterilization?

= to avoide class breakage (due to possible micro-cracks) & to guarantee product quality (no cracks, no residues in product, risk of contamination)

a) How is sterility defined according to Ph. Eur.?

= absence of viable micro-organisms

=> defined by a sterility assurance level (SAL) equal to or less than 10^−6

b) Two sterility test types

1) Membrane filtration

2) Direct inoculation

c) Minimum number of containers - eye drops 180 batch

=> more than 100 but not more than 500 = min. 10 containers

Because eye drops are usually in single-dose containers

How do the z-values for dry heat differ from those for steam sterilisation?

dry heat = 160° for 125 min

steam sterilization = 121.1°C for 1 min

Effectiveness = dry heat is less effective than steam sterilization at the same temperature because no condensation!

because:

1) microorganisms/ spores are more heat resistant under dry conditions (= D-values are approx. 10 times higher at the same temperature) => z-value 10°C

2) dry air has a lower heat capacity & poorer heat transfer and a higher risk of cold islands

= z-values about twice as high compared to steam sterilization because higher D-value necessary -> less effective/ no condensation

- FH value (instead of F0) - for dry heat sterilization

(a) Why are the vacuum-nitrogen cycles required?

ETO = highly effective sterilant but extremely flammable, forms explosive mixtures with air (O2) & toxic/ hazardous to human

• before sterilization:

  -> therefore O2 needs to be brought out of chamber by vacuum & for gas sterilization is nitrogen used

  -> through pulsation vacuumed input of nitrogen air gets better removed (better displacement)

• after sterilization: ETO = is toxic and may be potentially hazardous to humans. After the sterilization process is completed, it is crucial to remove residual ethylene oxide gas from the sterilized items to ensure they are safe for use. -> with vacuum ETO residues get sucked out & nitrogen (inert gas) gets introduced in chamber displacing ETO

(b) What is the purpose of the dwell period?

dwell-period = Sterilization time = exposure time of product under constant pressure / temperature & desirend conditions with sterilization agent (e.g. ETO)

- enables efficient penetration/ contact/ uniform distribution of sterlization agent over product

(c) Name one advantage of ethylene oxide sterilization over hydrogen peroxide sterilization.

Advantages - ETO = Excellent penetration capacity (highly effective)

(a) What steps are needed to biodecontaminate an isolator with hydrogen peroxide?

VHP = vaporized hydrogen peroxide = surface sterilization process / used for biodecontamination -> decrease in 10log levels

(b) Why should condensation of H2O2 be prevented if possible?

1) because drops on surface of product increase corrosion

2) duration process takes much longer if H2O2 gets to vapor

How would you evaluate the process of preparing and transferring the format part: Are all steps (1) to (5) in order with regard to the requirements of Annex 1 and a possible risk to the medicinal product in Class A? Is there potential for optimization? If yes, which one? (2 points)

1) wahing machine from class D to C = is right

2) Packed in a Tyvek-bad = not ok = the used of 2 Tyvek gas is needed to guarantee sterility

3) not ok = Handling & filling of aseptically prepared products in class A with class B environment

4) is right = create class A conditions

5) is right

(a) During the DQ you have to select a steam sterilization process for the sterilization of 2 m long silicone tubes used for aseptic filling. Which process do you choose? Give a justification for your choice.

DQ = Design Qualification = Determination of requirements for the equipment ->(URS = user requirements specifications & functional specification document)

Justification:

• with SIP is effectively sterilization of long tubing systems possible

• tubes can remain in their installed positions within the system during sterilization, maintaining aseptic integrity

• SIP systems are designed to ensure uniform distribution of steam, which is critical for sterilizing long silicone tubes uniformly. This prevents any cold spots or areas where steam might not reach effectively.

(b) What is the name of the document in which the user demands on a sterilization system are recorded as part of the DQ?

URS = user requirements specifications & functional specification document

(c) What is checked during thermoelectric testing with an empty sterilization chamber as part of the PQ of a steam sterilization process?

(a) Describe the venting process of a steam sterilization process using the fractionated vacuum method.

II) fractioned pre-vacuum process

= for special products with very difficult air elimination

- replay change of vacuum phase & steam introduction

- steam gets introduced to reach desired °C

- fractionated cooling & drying: vacuum & air

(b) How is container bursting prevented in the hot water spray process?

= through creation of counterpressure by introducing sterile compressed air into container

a) bobble point & pore diameter

bubble point = pressure at which a continuous discharge of air bubbles can be detected

=> the smaller the pore size = the higher the pressure (3-5 bar)

= the higher the bubble point

b) Give two reasons why an additional germ-reducing filtration is carried out.

bioburden reduction prefilters & sterilising grade filters

1) to prevent built up of the filter cake

2) if 1-st filter is having a leak the 2-nd filter enables filtration = increase safety of sterility

( 3) separation in 2 steps possible: one in class C area & one in final step)

What is the so-called bubble point of a sterile filter and how is it determined?

1) Bubble point test (manual application)

  = Membrane gets wetted with liquid product & pressure (air) gets applied from one side of the filter to press out the solvents out through the filter

-> Solvent is pressed out of filter with increasing pressure

Bubble Point = pressure at which a continuous discharge of air bubbles can be detected (3 to 5 bar) => depends on the filter properties

- big leak = lower pressure

- the higher pressure = the smaller the pore size

(a) List four different non-economic requirements that should be included in a user requirement specification (URS) for an autoclave

URS = user requirements specifications & functional specification document

= a planning document that specifies what a system needs to do => part of DQ (Design Qualification)

1) Location / Environment / Dimensioning

·   Place of installation/ Environment / zone requirements

·   Constructional conditions, e.g. available area, floor load

2) Process

·    Procedure / operating mode (e.g. FRVV)

·    Special requirements for the process (e.g. accelerated cooling)

·    Design / Type, (cylindrical/rectangular; horizontal/vertical)

·    Working range (pressure / temperature)/ Programms

3) Monitoring / Measurement technology

·    Sensors/ Accuracy/  Additional features

4) Materials and surfaces

·        Materials

·        Surface properties

·        Hygienic Design

(b) What must be considered in the PQ of a steam sterilization process with regard to loading - in addition to the loading schemes of routine production?

5) PQ = Performance Qualification = Verification of sterilization success based on relevant loads -> actual process validation

• PQ of a steam sterilization process = involves demonstrating that the sterilizer operates effectively and consistently within the defined parameters to achieve the desired level of sterility.

1) load & packaging system should contain product & be identical to the packaging used for routine production

2) Worst-Case Loading & Conditions should be included (max/ min/ cold spots)

3) loading configuration/ amount/ distribution inside (spacing) must be defined

(b) In preparation for the aseptic filling of a liquid medicinal product, the drug is usually subjected to two filtration steps. The second step is sterile filtration. What is the purpose of the first filtration step?

bioburden reduction prefilters and/or sterilising grade filters may be used at multiple points during manufacturing process to ensure a low & controlled bioburden before getting to the final sterilising filter

Purposes:

(c) Sterile filters must be examined for leachables. What are leachables?

Leachables (if required) = only required if toxic extractables detected

= Substances leached from the filter material in contact with the product under standard process conditions

= substances that can migrate or leach from materials used in the manufacturing process into the final product

-> for sterile filters, leachables refer to any chemicals or substances that may be released from the filter material and contaminate the filtered product.

(a) Which type of electromagnetic radiation used for sterilization has the highest frequency?

= gamma irradiation ( gamma rays)

Electromagnetic spectrum

b) Name one disadvantage of electron irradiation over gamma irradiation.

24) What is the advantage of electron radiation over gamma radiation?

= no radioactive waste & high dosage rate

(c) In which unit do you specify the dose for radiation sterilization?

(a) How are the F-value and the D-value related?

F-value [mins] = holding time at a temperature T in minutes to reduce the existing population (N0) with the specific D value by the desired number of powers often/ log10 levels (n) to a final value (N)

D-value = duration of sterilization to kill 90% of germs (of N0)

F-value is directly proportional to D-value.

• low F-value = short sterilization (holding time) => means low D-value = short duration of sterilisation to kill 90% of germs = steep curve

• high F-value = long sterilization (holding time) => means huge D-value = long duration of sterilisation to kill 90% of germs = flat curve

• The higher the F value, the … more extensive (better) is the killing of germs.

• The lower the F value, the slower is the SAL value achieved (low assurance of sterility)

• With increasing temperature the D-value [mins]… decreases -> high °C results faster killing & a less D-value

(b) For a given z-value of 10 °C, how does the D-value change if steam sterilisation is carried out at 120 °C (increased pressure, e.g. pressure cooker) instead of 100 °C (atmospheric pressure)?

= D-value decreases for factor 10^-2

(c) For a steam sterilisation process, the F0 value is 30 min. How can this value be interpreted?

F0 = 1 min means the lethality of a method that works for 1 minute at 121.1 °C (250 °F) and a z-value of 10 °C (18 °F) or an equivalent temperature-time combination

F0 value of 30min = means the same lethality of a method that works for 1 minute at 121.1°C

b) What should be considered when validating a steam sterilisation process with regard to loading?

1. Loading Patterns

• Standard and Worst-Case Loads: Validate using both routine (standard) and worst-case load configurations. Worst-case scenarios may include maximum and minimum load capacities, as well as challenging load items that are difficult to sterilize.

• Uniformity & Spacing : Ensure that the load is uniformly distributed within the sterilizer chamber to allow for proper steam penetration and heat distribution. Maintain enough spacing between items and ensure proper orientation to facilitate steam access to all surfaces and areas of the load.

• PQ = Performance Qualification = Verification of sterilization success based on relevant loads -> actual process validation

  • test load of the sterilizer contains products which are routinely handled -> loading patterns must be defined

  • load should represent product families which have the greatest difficulty for the sterilisation -> include worst case conditions

  • the loading configuration & amount of load must be defined Ò size and/or the mass of the sterilizer load

2. Material and Container Types

• Diverse Materials: Include a variety of materials (e.g., metal, glass, plastic, textiles) that are commonly processed to ensure that the sterilization process is effective for all types.

• Container Sizes and Types: Validate using different container sizes and types (e.g., vials, bottles, pouches, trays) to ensure steam penetration and effectiveness across all configurations.

3. Temperature and Pressure Monitoring

• Sensor Placement: Strategically place temperature and pressure sensors (thermocouples) throughout the load and chamber, especially in hard-to-sterilize areas, to ensure uniform conditions.

• Environmental Mapping: Conduct temperature and pressure mapping studies to identify potential cold spots or areas where steam penetration might be insufficient.

4. Biological and Chemical Indicators

• Biological Indicators (BIs): Place BIs in the most challenging locations within the load to provide direct evidence of effective sterilization. BIs should contain highly resistant microorganisms (e.g., Geobacillus stearothermophilus) to validate the process.

• Chemical Indicators (CIs): Use CIs as visual markers to confirm that sterilization parameters (e.g., temperature, steam penetration) have been met throughout the load.

5. Process Controls

• Cycle Parameters: Validate all critical cycle parameters, including temperature, pressure, and exposure time, to ensure they consistently achieve the required conditions.

• Steam Quality: Verify that the steam used meets the required quality specifications, including purity and dryness fraction, to ensure effective sterilization.

6. Repeatability and Reproducibility

• Multiple Runs: Conduct multiple validation runs to demonstrate that the sterilization process is reproducible and consistently achieves the desired outcomes under varying load conditions.

• Statistical Analysis: Analyze the data statistically to ensure consistency and reliability across all validation runs.

8. Regulatory and Compliance Requirements

• Compliance Standards: Ensure the validation process meets relevant regulatory standards and guidelines, such as those provided by the FDA, EMA, ISO, and other pertinent bodies.

• Documentation: Maintain thorough and accurate documentation of all validation activities, including protocols, test results, and analyses, to provide evidence of compliance and process control.

9. Load Configuration Specifics

• Dense and Light Loads: Validate both densely packed and lightly packed loads to ensure steam can penetrate effectively regardless of load density.

• Mixed Loads: Include mixed loads with different item types and materials to validate the versatility of the sterilization process.

c) What is the general name of test strips with bacterial spores used to validate sterilization procedures?

The general name for test strips with bacterial spores used to validate sterilization procedures is "biological indicators" or "biological indicator strips”

• biological indicator (for validation) = filter paper strips with spores/ spore suspensions with colour indicator

These strips contain a known number of highly resistant bacterial spores, such as Bacillus subtilis or Geobacillus stearothermophilus. Biological indicators are employed to assess the efficacy of sterilization processes, for steam (autoclaving), ethylene oxide gas, hydrogen peroxide gas etc.

(a) When may sterile filtration be used to sterilize a liquid medicinal product?

= if product is not heat stable at all, use sterile filtration

= if steam sterilization is not possible then sterile filtration with microbial retentive filter is the last option

=> must be combined with aseptic processing

 with presterilized containers -> only final sterile filtration & filling is done in class A

(b) As part of the validation of a sterile filtration process the bacterial challengetest (BCT) must be performed. How does it work?

Note: the viability test does not need to be addressed.

Bacterial Challenge Test = BCT – biological test for filtration

1) Preparation: performance of Viability test

2) Provide filtration process = under worst case conditions (with product, filter elements, contact time, pressure, temperature)-> check the filter retention capacity = validation of retention of germs on membrane filters

3) Checking sterility after filtration = 100 % retention of germs (acceptance criterion) -> no single cell of reference germ should pass the filter!

F0 & D-value

a) minimum SAL 10^-6 = 12 log levels

F-value = holding time (process time) to reduce N0 (with specific D-vlaue) by desired log-levels to final value N

(a) How can you draw conclusions about the integrity of a sterile filter from the measured value of the bubble point?

- big leak = lower pressure

- the higher the bubble point / pressure = the smaller the pore size

- The smaller the average pore size of a filter, the higher the pressure. If there is a leak in the filter the pressure (bubble point) stays low

(b) How is possible product loss during sterile filtration investigated?

with Adsorption test = for sterile filterable products

= checking the amount of product loss; muss be 100% (no loss)

= measure concentration of API before & after filtration

-> calculation of recovery = bust be 100% no change in concentration

a) Why must sterility testing be carried out in an aseptic process environment?

= to ensure the safety of aseptic process & reliability of the results

= to prevent the introduction of contaminants during the process

= because sterility must be verified for each batch to get released

b) Why can the direct inoculation method not be used for sterility testing of a sterile antibiotic?

sterile antibiotic = inhibits the growth of germs (function of antibiotic)

-> if product inhibits the growth of microbes = direct inoculation cannot be used because its not possible to do Positive method suitability test ( = to prove that the product doesn’t inhibit the growth) -> antibiotic does inhibit growth!

-> other sterility test must be applied

c) 2.4 mL => 1-40mL -> half the contents of ech container but not less tha 1 mL

-> half the content 2.4/ 2 = 1,2 => min. 1ml per container must be tested

(a) What would be the consequence if the product formulation was not carried out in at least class C, as required, but in class D?

Provide an explanation.

1) higher risk of product contamination of particles out of air or materials (“dirty” environment)

2) Impact on product quality -> may have unwanted particles -> danger for patient

3) Risk of bad sterility -> not sterile enough, to high bioburden, bad filtration -> no release of product (additional costs)

b) What must be checked on each filter before and after use?

= The integrity of the filter must be checked before & after use

-> after usage = easy to perform -> filter can get taken out for a quality control in lab or in production area

-> before usage = is critical because it must be assured that filter doesn’t get contaminated

 2 possibilities to keep filter sterile before usage:

1) sterilization in autoclave after filter testing Ò validation important

2) filter gets installed in production area & gets sterilized in place Ò filter testing can be done in place

(a) What can be criticized about the loading of this hot air sterilization oven?

• load should be putted in right position in the autolave (straight)

• product shouldn't stand on top of each other (air cannot get inside)

• air distribution = Air must be able to circulate freely

  -> metal sterilization cassettes with holes = product should be placed correctly on the trays (Tablett)

(b) For depyrogenation, hot air sterilization tunnels must be operated at over 220 °C according to the pharmacopoeia. The z value for depyrogenation is around 50 °C. What does this mean when you compare the required process time at 320 °C and 220 °C?

z-value =Temperature change in °C, which is required to alter the D value by a factor of 10 -> 1 log reduction

= process time is longer at 220°C

= process time at 320°C is reduced by factor 100 compared to the time at 220°C

(15min = 900sek -> 900sek/9sek = 100)

Describe all further steps or processes required until sterilization is complete. Also address the issue of air removal.

2) water heats up -> generation of steam (with gas burner)

3) stean flows inside the vessel

4) steam rises up, reaching the lid -> steam moves down to sterilisation items

-> product is cold -> steam condensates

-> air removal = steam presses the air downwards & out of the system (via air separator, open valve)

5) atmopsheric pressure = air is eliminated & temperature can rise to 121 °C -> valve closes when all air is out -> pressure inside rises to achiieve p= 2 bar

6) Sterilisation gets done for desired time ( at least 15 mins) at 121°C

7) Cooling dwn of system -> until temperature below 80°C

a) SAL value of 1.4 * 10^-6 is NOT achieved. = 1.4 container of 1 mio non-sterile

EMA - specifiation: 1* 10^-6 = max 1 container of 1 mio non sterile

= the process doesn’t meet the specification of EMA because 1.4 is higher than 1

b) cfu/ml = 1

1 - log (10) = 0

Probability of sterility = 100%

a) = 131,5

b) How is the achieved F0 value of 50 min to be interpreted?

the F0​ -value of 50 minutes = SAME lethality of germs that is achieved for 50 minutes at 121.1 °C

(a) How do you call the release of terminally sterilized products without batch-related sterility testing, based solely on the measurement of physical process parameters?

b) What is checked during the OQ of a steam sterilizer?

OQ = Operational Qualification

  = Checking basic functions of equipment within the working area including chamber integrity (pressure & vacuum test)

(c) What is the acceptance criterion for the Bacterial Challenge Test (BCT)?

Checking sterility after filtration = 100 % retention of germs (acceptance criterion) -> no single cell of reference germ should pass the filter!

a) For which sterilization goods is the steam-air mixture process used?

1) steam/air mixture process

(b) Why does the steam-air mixture process use a steam-air mixture and not pure saturated steam?

 = because to generate steam higher °C is needed, which raises the pressure: p = p1 + p2 -> container may burst

-> higher °C to create steam may have negativ impact on product

Reasons for using a steam-air mixture:

I) Temperature Control:

• Reduced Temperature: Pure saturated steam provides uniform heating at fixed temperature corresponding to its pressure (e.g., 121.1°C at 1 atm). But for sterilization processes with lower or more precised temperatures stea,-air mixtures are necessary.

• Mixture Adjustments: By mixing steam with air, the overall temperature can be adjusted below the saturation temperature of pure steam. This enables more controlled heating environment for delicate items or processes that cannot withstand higher temperatures.

II) Uniform Heating

• The addition of air can help in distributing heat more uniformly

• Avoiding Condensation Issues: Pure steam can condense on cooler surfaces, leading to uneven heating and potential cold spots. A steam-air mixture can reduce the rate of condensation, leading to a more uniform temperature profile within the chamber.

• steam-air mixture allows better control over temperature. Pure saturated steam can have a high temperature & cause damage on products. By introducing air into the steam, the overall temperature can be moderated to prevent overheating

(a) Why does the formulation of the product have to be carried out at least in clean room class C, when subsequently two filtration steps are carried out to achieve sterility?

= to ensure a low & controlled bioburden before getting to the filtration

-> according to EMA-guidline = Frequent bioburden limit: ≤ 10 CFU / 100 ml -> to fulfill this clean room conditions necessary

= the lower the bioburden, the slower cake formation during filtration (during dead-end filtration)

(b) What must be checked on each filter before and after use?

= Filter integrity testing– GMP Guidline

= The integrity of the filter must be checked before & after use

-> after usage = easy to perform -> filter can get taken out for a quality control in lab or in production area

-> before usage = is critical because it must be assured that filter doesn’t get contaminated -> 2 possibilities to keep filter sterile:

1) sterilization in autoclave after filter testing -> validation important

2) filter gets installed in production area & gets sterilized in place -> filter testing can be done in place

(a) A low bioburden before sterilization is not only important to achieve the shortest possible sterilization time and the highest possible sterility assurance. What other reason is there?

• Prevention of undesirable by-Products -> metabolic-products of germs may begatively influence the quality of product

• low bioburden is advantageous for sterile filtration

  -> filter capacity = with high bioburden gets quickly blocked by germs (cake formation)

  -> depending on amount of bioburden the filter must be huge enough

(b) How are the D-value and the z-value related?

relationship in formula:

z-value = Temperature change in °C, which is required to alter the D value by a factor of 10 -> temperature dependency of D-value

9)

It means that the steam sterilization process for 30min at 125°C reaches an equal lethality as the process for 74 mins at 121.1°C.

(a) What is the minimum cleanroom class in which the preparation must take place?

= Class C

b) Give two reasons why additional germ-reducing filtration is carried out.

bioburden reduction prefilters and sterilising grade filters

1) to prevent the build up of filter cake, first filter takes the main germs load out of product

2) if the first filter is having a lack the second filter enables the filtration (safety of process)

3) increases the process safety to ensure a sterile product

4) separation of the 2 steps is possible, so one is done in class C area & the other in the final filtration

Why, in critical cleanroom classes, cleanroom personnel should make only slow, controlled movements?

• at high clean room classes are high requirement for aseptic working

• slow controlled movements to avoid:

  1) interruption of unidirectional flow or creation of air tubulances

  2) contamination of products or spread out additional particles

How do you evaluate the process?

= the process doesn’t fulfill the criteria of aseptic preparation

Are all steps (1 to 5) in compliance with the requirements of annex 1 and a minimal contamination risk for the drug processed in class A?

= No are not, high contamination risk due to wrong unpacking in wrong zone

Is there potential for optimization? If yes, which?

[1] ist right

[2] & [3] = tubs get unpacked/ opened (removal of second packaging tyvek bag) in class C -> surface sterilization from C to B is correct

[4] door of RABS shouldn't get opened -> high contamination risk (class A inside) & unpackaging of second packaging layer has to be in class C

[5] doors shouldn't open (transporttation via decontamiantion airlock or RTP better) - inner tyvek bag better to open in class B area & transfer sterile tub to class A for final filling/ processing

Main steps:

I) Class C/D: 1) tubs get mechanically unpacked/ opened -> HEPA filtered air

 2) tubs get introduced into E-beam tunnel -> through airlock doors

 3) tubs get transferred to irradiation zone -> surface sterilisation

II) Class B: 4) tubs get transported trough class B zone to class A

III) Class A: 5) filling zone -> presterilized syringes/ vials get filled

State two principles of aseptic work. What the consequences of not following the principle would be for the production environment or process.

Principles of aseptic working

1)  Do not work above the product = risk of product contamination & risk for inhalation of (toxic/ bad) substances for personnel

2) Desinfect sterile gloves regularly or change them = to avoid spread out of particles, parts of skin (sweat) getting to product or particels from product getting through the gloves (protection of personnel)

What is the leak test procedure for an isolator glove?

2) Leak testing:

 2.1) Pressure decay test = gloves get blowed up with pressure

 2.2) airflow test at constant pressure = air gets inside of gloves & measurement of volume that is needed to keep pressure constant -> if tight, no additional air needed

(a) What is measured to monitor CIP processes?

= is a Validated automatic cleaning without disassembly

-> Mobile or fixed systems with connection to SIP

Monitoring = measurement of Conductivity (possibly also pH / TOC = total org. carbon) in the effluent (Abwasser)

Conductivity = property of water to conduct electrical current. Is influenced by the concentration of ions in the water. => used as an indicator of overall water quality.

High conductivity = high concentration of dissolved ions in water

= may suggest the presence of contaminants or changes in the composition of water.

(b) Sterile couplings are used for single-use systems. What is their purpose?

Sterile connectors/ couplings = presterilized system with membraned (gets taken out) for product transfer

= for Hose connection with sterile couplings (under class A air) = disposable system for product transfer in class A to maintain sterility

- tiny hole in the wall between class C & class A (overpressure)

- tube can be fixed in this hole & get connected with sterile coupling

-> reduced risk of recontamination

(c) In addition to the glass body, which parts of a finished syringe are also siliconized?

= syringe stopper & needle (if present)

(a) Which two transfer technologies are best suited for transferring crimping pliers into an isolator during operation?

1) RTP = Rapid Transfer Port = double door transfer systems

 = dog removable storage/ containers bag -> not directly connected to isolator, connection via alpha port

= for transport of stoppers, caps & Liquids

2) Decontamination airlock

(b) E-beam tunnels: How is the ingress of germs from the loading area (insertion of the still film-wrapped tubs) into the filling area (Class A) prevented?

= through pressure cascade & physical barrier (walls)

(a) From what can it be determined that the RABS shown is an open, active RABS?

open = air circulation in & outside chamber (from A to B class)

active = HEPA Filter inside

-> independent air supply unit

(b) Which GMP clean room class is required as the background environment if aseptic production takes place in this RABS?

= class B environment (for RABS)

c) What is the difference between RABS & Isolator?

a) open active RABS

b) differences between a product protection isolator and a personal protection isolator (for handling a non-sterile toxic product).

• product protection isolator

  1) protection of product -> operation at overpressure +

  2) working with gloves

  3) Fully sterilizable (H2O2 decontamination)

  
  

• personal protection isolator

  1) protection of personnel

  - Negative pressure = operation for non-aseptic products

  + Overpressure = for aseptic/ toxic products

  
  

  2) working with special manipulators (NO gloves!)

  3) Spray cleaning (CIP) = manual spray cleaning from outside by personnel

  4) exhaust air protected = air gets filtered bevor leaving isolator

(a) What characterizes an open, passive RABS?

(b) What characterizes an open isolator?

II) Open isolator systems = allows continuous material flow, hermetic seal for decontamination, with unidirectional air flow

 = designed for continuous or semi-continuous ingress and/or egress of materials (Ein-/ Austritt) during operations through one or more openings -> mouse holes

 = Openings must be engineered (e.g. using continuous overpressure) that no external air/ contaminant get into the isolator

-> has tiny openings (“mouse holes”) with inside overpressure to ensure that no particles get inside

-> min. grade C surrounding (because of holes)

(c) When is underpressure used within isolators?

- Negative pressure = operation for toxic/ hazardous products

(d) How often / at what frequency should the leak test of an isolator glove be carried out?

= MUST be done at each batch change before every biodecontamination -> prove if isolator & gloves are tight/ closed

A beta container is to be used for the transfer. Describe the process for transferring the forceps

1) Rapid Transfer Port = RTP

= for transfer of materials, stoppers, caps into aseptic area while processing

Principle:

- double door transfer system

- consists of an Alpha-port which is installed in the wall of isolator or RABS

- for transfer a Beta-container/ bag gets connected to Alpha-port

 -> beta-container fits exactly to alpha-port

1) Beta container = stainless steel with pipe connector & hydrophobic sterile filter = forceps are put in the beta container

2) container (with forceps inside) gets autoclaved with vacuum

3) Connection of Isolator + container -> connected lids get opened from inside for material transfer = must be air-tight to avoid decontamination

-> lids must be airtight connected ->opening of lid only possible when fully connected-> by rotation 60°

4) forceps can be transfered into isolator & there used

-> higher pressure inside transfer pressure for product protection

(a) Which RABS type is shown in the figure below and what is the main difference to an isolator?

open = air circulation between zones (A to B)

active = HEPA filter Unit inside

= open, active RABS

• Main difference to Isolator = surrounding environment ISO 7 which is equivalent to class B -> Isolator has an surrounding environment of C/D

(b) How often / at what frequency should the leak test of an isolator glove be carried out?

= MUST be done at each batch change before every biodecontamination -> prove if isolator & gloves are tight/ closed

a) What checks/tests must be performed on the gloves before usage?

1) Visual inspection of the gloves -> if leak detected no further test needed

2) Leak testing:

 2.1) Pressure decay test = gloves get blowed up with pressure

 2.2) airflow test at constant pressure = air gets inside of gloves & measurement of volume that is needed to keep pressure constant -> if tight, no additional air needed

b) principle of a transfer into an isolator during operation using a decontamination airlock.

c) Indicate one way in which a continuous transfer of material can be made out of an isolator?

(a) What is the meaning of the abbreviation RABS?

= Restricted Access Barrier System

(b) What characterizes an active, closed RABS?

(c) When is overpressure used within isolators?

= always for aseptic purposes = + over-pressure inside (protection of product)

(a) Which medium is used in a CIP process for the final rinse of a format part that is later used in an aseptic process?

= WFI = Water for injection

(b) Name one way in which two silicone tubes can be aseptically connected.

= aseptic connection via:

Sterile welding of thermos plastic tubes = 2 tubes get brought together & closed in a special machine (welder) -> inside machine: tubes get cut & welded to each other -> class A air supply

(c) What process takes place for cartridges after washing before treatment in the hot air sterilization tunnel?

= siliconization

(d) What is the final process step of an automated stopper treatment?

Capping/ Crimping = after placing a stopper on a vial, capping must be perform to fully seal the vial

 = ONLY after this step container is considered ready & sealed -> can leave class A

(a) In CIP processes, it is important that no spray shadows occur. What is meant by this?

"spray shadows" = areas or surfaces within a processing system that are not effectively reached or cleaned by the spray nozzles during the CIP cycle.

(b) Stoppers that will later be used in syringes in an aseptic process are to be prepared. Outline the process steps of automatic stopper treatment (excluding transfer and storage processes).

Automatic stopper treatment

1) Washing & cleaning = to remove any contaminants, particles, or residues.

2) Rinsing = After cleaning, the stoppers undergo rinsing to eliminate any remaining cleaning agents or impurities

3) Drying = to remove moisture

4) Sterilization = autoclaving, gamma irradiation

5) Siliconization = application of a thin layer of silicone oil to the stopper surfaces. This is done to facilitate the movement of the stoppers within the vial or syrange and reduce frction & enebale correct dosing

6) Inspection and Quality Control = to ensure that quality: visual inspection for defects, dimensional checks, and testing for sterility.

7) insert stopper in syringe (with pipe or vacuum)