The Main Sources of Contamination in the Aseptic area (Pharmaceutical Industry)

Contaminants can gain entry into a production process stream from several sources such as, Personnel,
Poor facility design, Incoming ventilation air, Machinery and other equipment for production, Raw
material and semi-finished material, Packaging material, Utilities, Different media used in the
production process as well as for cleaning and Cleanroom clothing. Let’s learn about these sources in
detail.

1. Personnel
 Personnel who are supervising or performing drug manufacturing or control can be a potential
source of microbiological contamination and a vector for other contaminants. The main reasons
for contamination from the personnel include:
 Lack of training
 Direct contact between the operator’s hands and starting materials, primary packaging
materials and intermediate or bulk product
 Inadequate personnel cleanliness
 Access of unauthorized personnel into production, storage, and product control areas
 Inadequate gowning and personnel protective equipment, and
 Malpractices like eating food, drinking beverages, or using tobacco in the storage and
processing areas.

2. Buildings and Facility

 The buildings and manufacturing facilities may also contribute to the contamination. The main
reasons of contamination due to facility issues include:
 Insufficient size and inadequate organization of the space leading to selection errors like
mix-ups or cross contamination between consumables, raw materials, in-process materials,
and finished products
 Inadequate dirt and pest controls
 Rough floors, walls, and ceilings
 Lack of air filtration systems Improper lighting and ventilation
 Poorly located vents, ledges, and drains, and
 Inadequate washing, cleaning, toilet, and locker facilities to allow for sanitary operation,
cleaning of facilities, equipment, and utensils; and personal cleanliness

3. Equipment
 The equipment and utensils used in processing, holding, transferring and packaging are the
common source of pharmaceutical contamination. The main reasons for contamination from
the equipment include:
  Inappropriate design, size, material leading to corrosion and accumulation of static material
and/or adulteration with lubricants, coolants, dirt, and sanitizing agents
 Improper cleaning and sanitization
 Design preventing proper cleaning and maintenance
 Improper calibration and irregular service, and
 Deliberate use of defective equipment

4. Materials
 The raw materials used for production can be a potential source of contamination. The main
reasons for contamination from the raw materials include:
 Storage and handling mistakes causing mix-ups or selection errors
 Contamination with microorganisms or other chemicals
 Degradation from exposure to excessive environmental conditions such as heat, cold, sunlight,
moisture, etc.
 Improper labelling
 Improper sampling and testing, and
 Use of materials that fail to meet acceptance specifications.

5. Manufacturing Process
 There are various opportunities for contamination of raw material, intermediates or packaging
materials throughout the manufacturing process.
 The main reasons for contamination during manufacturing process include:
 Lack of dedicated facilities to manufacture a single product
 Inappropriate cleaning in-between batches to minimize the amount of product changeovers
 Use of an open manufacturing system exposing the product to the immediate room
environment
 Inappropriate zoning (divide into or assign to zones)
 Absence of an area line clearance according to approved procedures following each
cleaning process and between each batch, and
 Lack of cleaning status labelling on all equipment and materials used within the
manufacturing facility

 To minimize the risks of manufacturing contamination

 Manufacture products in a campaign, with the appropriately qualified cleaning processes and
checks performed in-between batches to minimize the amount of product changeovers
 Utilize a closed manufacturing system.
 This is where the product is not exposed to the immediate room environment (and vice versa)
  Perform an area line clearance according to approved procedures following each cleaning
process and between each batch/campaign Zone the facility and
 Use Cleaning Status labelling on all equipment and materials used within the manufacturing
facility

6. HVAC System
 A poor HVAC system can be a potential source of microbe’s growth and a transportation mode
for dispersing contaminants throughout the manufacturing facility.
 The main reasons of contamination due to HVAC issues include:
 Accumulations of organic material in or near HVAC air intakes
 Ineffective filtration of the supply air
 Insufficient magnitude of pressure differentials causing flow of reversal
 Erroneous ratio of fresh air to recirculated air
 Inability to access ventilation dampers and filters from outside the manufacturing areas, and
 Non-directional airflow within production or primary packing areas

Establishing a Contamination Control Strategy for Aseptic Processing

 Sterility is a key quality attribute for a class of medicines required to be sterile. The
consequences of non-sterility are direct patient harm. The likely outcomes of the
administration of a non-sterile product are disability or death.
 For this reason contamination control is of utmost importance for the manufacture of sterile
products and this is especially important for products that are filled aseptically, where terminal
sterilization is not possible.
 Aseptic manufacture involves the production of drug products which are not subject to a
sterilizing step; instead, sterility is assured through the prevention of microbial ingress.
 To assess the risks of non-sterility each organization should develop a contamination control
strategy. This requires an assessment, acknowledgement and remediation process for
contamination risks.
 The objective of a contamination control strategy, for aseptically filled products, is sterility
assurance and the production of a sterile product.
 Aseptic Processing: The process of producing a sterile product by aseptic processing is either
through conventional filling or by blow-fill-seal. With both, a product is sterile filtered into a
sterile container and filled into depyrogenated containers and then sealed. The bringing
together of the sterile product and the container is performed under ISO 14644:2015 Class 5
conditions.
1. Good Facility Design
 All sterile product manufacturing must, according to regulations, be undertaken within a
classified cleanroom environment. This is in order to minimize product contamination.
 Shortest product flow paths possible to ensure there are airlocks in place between cleanrooms
of different grades and for changing rooms.
 Good design relates to both selecting a suitable grade of cleanroom together with a design
intended to minimize contamination. This includes the use of appropriate construction
materials and spending time on the suitability of the layout, covering elements like process and
material flows.
 With this the layout should promote the orderly handling of materials and equipment, the
avoidance of mix-ups, and the prevention of contamination of equipment or product by
chemical substances, previously manufactured products, and microorganisms.
 Cleanroom must have suitable air change rates, air mixing and be able to recovery rapidly after
a contamination event.

2. Personnel Training and Gowning

 With people representing the primary source of contamination effective training of operators is
paramount.
 Operator training should be continuous, covering theoretical, practical and cGMP aspects; with
the curriculum including microbiology and hygiene.
 The suitability of gowning should be assessed through regular gown qualifications, which need
to be assessed both visually and through microbiological sampling.

3. Cleaning and Disinfection

 Regular cleaning (using a detergent to remove soil) and disinfection (to inactivate
microorganisms through cellular destruction) is required.
 It is typical to use two disinfectants on rotation, one of which is often a sporicide (capable of
destroying bacterial endospores and fungal spores).
 For batch filling, cleaning and disinfection of cleanrooms must take place before and after each
run (and also of the conventional filling area should a closed RABS or isolator not be used).

4. Robust Sterilization
 The use of sterile items, from vessels to closures, needs to be practiced and controlled.
 Increasingly such items are of a bespoke design and are introduced as sterile and ready-to-use;
however, there remains a strong reliance upon in-house sterilization and this is most commonly
undertaken using an autoclave.
  To assess the effectiveness of different autoclave loads, as used with different cycles, these
need to be evaluated thermometrically and using biological indicators prepared from
Geobacillus stearothermophilus.

5. Use of Single-Use Sterile Disposable Items

 The use of single-use sterile disposable items has helped to move aseptic processing forwards,
through reducing the reliance upon autoclaves and helping to guard against both a failure with
a sterilization cycle and a control breakdown when a critical path step is undertaken, such as an
aseptic connection.
 Types of single-use technologies relevant to aseptic processing include tubing, capsule filters,
single-use ion exchange membrane chromatography devices, single-use mixers, and
bioreactors, product holding sterile bags, etc.

6. Depyrogenation
 Concern with pyrogens in aseptically prepared products (especially bacterial endotoxin in
relation to parenteral products) requires that the material into which the sterile bulk is
dispensed is depyrogenated.
 For glass vials this is via a depyrogenation tunnel (typically dry heat).
 Depyrogenation of glassware is important in the production of parenteral pharmaceuticals as
residual pyrogens could ultimately be injected into a patient resulting in an adverse reaction.
 The assessment of depyrogenation involves a formal study using thermometric measurements
and the use of endotoxin indicators.

7. Aseptic Filling and Barrier Technology

 With aseptic filling the sterile product is filled into depyrogenated glassware and fitted with a
sterile stopper and then over sealed.
 The most vulnerable step is with the dispensing of the product, via filling needle, into the vial.
Regulations require this to be undertaken in ISO 14644-1:2015 Class 5 conditions (with particle
and microbial control).
 With these design space choices there is a cascade of control in terms of automation and
consistency of the decontamination process; and with the barrier between the critical area and
the outside environment. Here the isolator provides a complete barrier.
 Leakage needs to be assessed, of both the isolator system and of glove ports.
 In addition, the airflow within the isolator (as with any aseptic filling zone) needs to be visually
assessed, using smoke or fog, in terms of having a suitable velocity and direction so that any
contamination that might gravitate towards a critical area like point of fill is directed away.
8. Media Simulation Trials
 The media simulation trial provides the means to challenge the aseptic processing assurance
system.
 With media simulation trials, a microbiological growth medium is used in place of the product
and filled as if it was product under the ordinarily processed conditions.
 Media fills start with the beginning of filling operations, during and after manipulations and
interventions, and until the last vial has been filled.