Aseptic Techniques Mfg Ii

Prof.: Jacqueline Hernández, M.S.

Module Outline Scope  Regulatory Requirements  Aseptic Processing  Microcontamination  Types of Contamination  Source of Contamination  Personnel 

Module Outline  Equipment and Facilities  Production Material and Processes  Improper Work Practices  Controlled Environmental Concepts ◦ Contamination Pathway ◦ Cleanroom Parameters

 Module

Outline  Personnel and Contamination Work Practices  Cleaning the Cleanroom ◦ Housekeeping  Microbiology

Laboratory

- Aseptic Techniques ◦ General Principles that should be observed

 CGMP’S

Part 211  Subpart C – Building and Facilities  Subpart D – Equipment  Subpart F - Production Controls and Manufacturing  Subpart I – Laboratory Controls



Regulatory Requirements:

◦ <1211> Sterilization and Sterility Assurance of Compendial Articles  Provides a review of the principles involved in producing aseptically processed products with a minimal risk of microbial contamination in the finished lot of final dosage forms.

 Regulatory

Requirements

◦ Chapter USP <1211> ◦ The areas of critical concern are the immediate microbial environment where these pre sterilized components are exposed during assembly to produce the finished dosage form and the aseptic filling operation.

 Regulatory

Requirements  Chapter USP <1211> ◦ The requirements for a properly designed, validated and maintenance filling or other aseptic processing facility are mainly directed to:  an air environmental free from viable microorganisms

 Regulatory

Requirements  Chapter USP<1211> ◦ Proper design to permit effective maintenance of air supply units ◦ Provision of trained operating personnel who are adequately equipped and gowned.

 Properly

designed should have the following considerations: ◦ Nonporous and smooth surface  Including walls and ceilings ◦ Gowning rooms with adequate space for personnel and storage of sterile garments. ◦ Adequate separation of preparatory rooms for personnel from final aseptic processing rooms

 Regulatory

Requirements <1211>

◦ Device as airlocks ◦ Air showers ◦ Proper pressure differentials between rooms ◦ Employment of laminar (unidirectional) airflow in the immediate vicinity of exposed product or components.

 Regulatory

Requirements

<1211> ◦ Filtered air exposure with adequate air change frequency. ◦ Appropriate humidity and temperature environmental controls ◦ Documented sanitization program

 Regulatory

Requirements <1211> ◦ Proper training of personnel in:  Hygienic Gowning techniques  Gowns Gloves Other body coverings substantially cover exposed skin surfaces.

 Regulatory

Requirements <1211> ◦ Certification and Validation of the aseptic process and facility. Efficiency of the filtration systems Employing microbiological environmental monitoring procedures. Processing of sterile culture medium as simulated product.

 Regulatory

Requirements <1211> ◦ Monitoring of the aseptic facility Periodic environmental filter examination Routine particulate examination Microbiological environmental monitoring Periodic sterile culture medium processing



Regulatory Requirements <1211>

◦Sterility Testing The facility should be such as to offer no greater a microbial challenge to the articles being tested than that of an aseptic processing production facility.

 Regulatory

Requirements <1211>

◦ Sterility Testing  The sterility testing procedure should be performed by individuals having a high level of aseptic techniques proficiency. The test performance records of these individuals should be documented.

 Clean

area control parameters should be supported by microbiological and particle data obtained during qualification studies.



It is important for area qualification and classification to place most emphasis on data generated under dynamic conditions ◦ Personnel present ◦ Equipment in place ◦ Operations ongoing



US Federal Standard 209D ◦ Establishes standard classes of air cleanliness for airborne particulate levels in clean rooms and clean zones. ◦ Clean room class in the statistically allowable number of particles, greater than or equal to 0.5 micrometer in size, per cubic feet of air.

 Definitions  Is

a room in which the concentration of airborne particles is controlled to specific limits.  They are classified by the numbers of particles per cubic foot per minutes.

 Sizes

of particles:

◦ which are controlled usually range from 0.1 micron to 10 microns.  Typical

room classes are Class 1, 10, 100, 1,000, 10,000 and 100,000.  Classes in ISO terms 1, 2, 3, 4, 5, 6, 7 and 8.

A

critical area is one in which the sterilized drug product, containers and closures are exposed to environmental conditions that must be designed to maintain product sterility (211.42(c)(10)

This area is critical because an exposed product is vulnerable to contamination and will not be subsequently sterilized in its immediate container.  To maintain product sterility, it is essential that the environment in which aseptic operations area conducted be controlled and maintained at an appropriate quality. 

One aspect of environmental quality is the particle content of the air.  Particles are significant because they can enter a product as an extraneous contaminant, 

◦ and can also contaminate it biologically by acting as a vehicle for microorganisms.



Class Limits in Particles per Cubic Feet

Room Class

0.1 0.2

0.3

0.5

5.0

1 10 100 1000 10,000 100,00

35 350 n/a n/a n/a n/a

3 30 300 n/a n/a n/a

1 10 100 1000 10,000 100,00

n/a n/a n/a 7 70 700

7.5 75 750 n/a n/a n/a

The nature of the activities conducted in a supporting clean area determines its classification.  FDA recommends that the area immediately adjacent to the aseptic processing line meet, at a minimum, Class 10,000 (ISO 7) standards under dynamic conditions. 

 Manufactures

can also classify this area as Class 1,000 (ISO 6).  An area classified at a Class 100,000 (ISO 8) air cleanliness level is appropriate for less critical activities ◦ Equipment cleaning

Contamination

Material which we do not want. There are a multitude of material and processes both biological and industrial, that are sensitive to contamination.

 An

essential part of contamination prevention is the adequate separation of areas of operation.  To maintain air quality, it is important to achieve a proper airflow from areas of higher cleanliness to adjacent less clean areas.

It is vital for rooms of higher air cleanliness to have a substantial positive pressure differential relative to adjacent rooms of lower air cleanliness.  For example, a positive pressure differential of at least 10-15 Pascals (Pa6) should be maintained between adjacent rooms differing classification (with doors closed). 

 When

doors are open, outward airflow should be sufficient to minimize ingress of contamination, and it is critical that the time a door can remain ajar be strictly controlled.

The Agency recommends that pressure differentials between cleanrooms be monitored continouosly throughout each shift and frequently recorded.  All alarms should be documented and deviations from established limits should be investigated 



Contamination It may be obvious to us when it is in the form of dust and lint on a work surface. It may be in the less distinguishable form of vapor in the air.

Types

of Contamination

 Organic  Inorganic Substances



Inorganic Materials  Can be natural  or Man made solids, liquids or gases They can be elemental = consisting of one elements of the periodic table. Compounds = chemical combinations of two or more elements (ex. Sodium chloride) Mixtures = such as metal alloys (ex. Brass)

Organic

Contaminants

 Complex chemical structures are typical of organic contaminants if they are of biological origin.  Three Categories:  Particulate Materials  Bacteria, fungi or other materials generated by biological processes  Gases and vapors



Particulate Materials Consist of metal fragments, lint from clothing, skin flakes and human hair. Clearly every effort must be made to reduce the number of these particles to an absolute minimum.



Bacteria, Fungi…  Bacteria and fungi can grow on a wide variety of material in light and darkness. The residue of these growth processes can be highly toxic or sufficiently alkaline to each material.  We can limit their rate of growth to the point where they do not present a severe problem.



Gases and Vapors

 Condensed organics can serve as growth media for bacteria or fungi.  Variety of materials can evaporate and then condensed as films or particles.



Airborne Particulates Contamination is usually in small quantities compared to whatever is being contaminated. Solid contamination typically will take the form of small particles in a range of sizes.



Airborne Particulates Visible particles are in the order of 50 microns. Bacteria range from 0.3 microns. Face powder is in the range 0.5 micron to 10 micron. Smoke ranges from 0.01 micron to 1 micron.

 Particles

may be transported to our sensitive site by: Air Water Process chemicals Gases On the surfaces of process solids Packaging Skin



Airborne Particulates Once particles are generated and released from their source materials, they may be carried by air currents generated by meteorological conditions: Wind Movement of people Movement of equipment Convection

Gravitational

force = mass x acceleration due to gravity The drag forces are not so easily represented. Depend on a number of factors such as the density and velocity of the air and the nature of the particle.

Airborne

Particulates

 Near-spherical particles greater than 1 micron in diameter will settle quite quickly under the force of gravity.  This is important will result in the removal of around 99.9% of these larger particles from still air.



Airborne Particulates  Particles smaller than 1 micron but larger than 0.1 micron in diameter will settle very slowly.  will be carried readily by even slight air movements.



Airborne Particulates

Particles less than 0.1 micron in diameter still totally in air. These particles will tend to remain suspended almost indefinitely.

 there are no moving air drag forces.



Airborne Particulate  The mechanisms here involves the motion of the gas molecules. In air at room temperature the gas molecules are moving with an average velocity in random directions.  Depend on temperature



Brownian Motion The particles “dance around” in the still air as they are hit from all sides by the molecules.



Gases and Vapors Gaseous contamination may also be transported in moving air by:  mixing  in still air by diffusion



Gases and Vapors Transport due to mixing will depend on the velocity and turbulence or the flow. Transport due to diffusion depends on the diffusion rate of the gas in air. Depends on the type of gas and the concentration gradients.



Gases and Vapors  Vapors may react from the vapor phase, but they may also condense as a thin layer of liquid or semisolid on the surface. Example: oil vapors may recondense on a cool surface to create a thin oil film.

Contamination

in Liquids

If particles are created of fall into a liquid, they may be readily transported by that liquid in the same way as air transport them. Liquids transport larger particles than air at the same velocity.

Contamination

in Liquids

A further problem with particles in liquids is that the particle material may be soluble to some extent in the liquid. Ex. Many minerals dissolve in water and some organic substances in alcohols.

Contamination

in Liquids

This disperse the contamination and makes its removal very difficult. Gases and vapors may also become dissolved in liquids. Unwanted liquid materials may also contaminate liquids Ex. Alcohol in water

Contamination

in Liquids

 Liquid contamination need not mix with the water to be transported.  Ex. Most oils do not mix water but will be carried with it.

Personnel

 One highly significant source of contamination internal to the production environment is the personnel who work in that environment.



Personnel  Personnel are critical for two very good reasons: Humans are animals and as such are biological contamination “factories” The operators are frequently the closest thing to the actual elements we wish to keep free from contamination.



Personnel “Dirty People” Cleanest people produce huge amounts of contamination as part of the process of living.

Personnel  The most significant contamination produced by humans is skin flakes.  The skin we are covered with is there to protect us from the environment we live in.  We are regularly bombarded by harmful ultraviolet rays which can actually destroy cells and tissue. 



Personnel The human body defends itself by sacrificing its outermost layer of skin and allowing it to become disposable.

The dead cells flakes off when the skin is gently abraded and will be carried off by: the convection currents surrounding the body or channeled through the clothing.



Personnel  The cells are organic  They are complex in a chemical sense  They carry other substances which are naturally found on the skin. Ex. Sodium chloride from perspiration  skin oils



Personnel

The number of skin flakes particles shed depends:  the state of the skin  the activity of the subject

Personnel

 If the skin is moistened by perspiration or by the application of a moisturizing skin cream, the skin flakes will tend to stick to the surface better.



Personnel

Body heat will result in the drying out of the skin and the subsequent release of the cells. Dry cells will also tend to fracture into many small particles.

Personnel 

Move Slowly and Deliberately ◦ Rapid movements can create unacceptable turbulence in a critical area. ◦ Such movements disrupt the unidirectional airflow, presenting a challenge beyond intended cleanroom design and control parameters.

Personnel  Move

Slowly and Deliberately (cont’)

◦ The principle of slow, careful movement should be followed throughout the cleanroom. ◦ Keep the entire body out the path of unidirectional airflow.

 Personnel  Move

Slowly and Deliberately (cont’) ◦ Unidirectional airflow design is used to protect sterile equipment surfaces, container closure and product. ◦ Disruption of the path of unidirectional flow air in the critical area can pose a risk to product sterility.

Personnel  Move

Slowly and Deliberately (cont’) ◦ Approach a necessary manipulation in a manner that does not compromise sterility of the product.



Personnel

A good rule of thumb is that a motionless person will generate some 100,000 particles of approximately 0.3 to 0.5 microns diameter per minute.



Personnel This figure changes dramatically when the subject begins an activity. As the increased abrasion of clothing against skin and limbs against body, As the constant expansion and contraction of the flexible skin will result in the expulsion of a greater number of particles.



Personnel A person sitting at a workbench moving just the arms and body generate on average 1,000,000 particles per minutes.

Personnel

If the whole body is in motion, for example during walking, this number increases by at least a factor of five.



Personnel Skin flakes are not the only contamination that the human body produces. Small flakes of the protein keratin are released from the hair. Hair itself will also be released from all over the body.



Personnel The act of breathing produces a great many contaminants.  In addition to large quantities of carbon dioxide and water vapor.  there will be droplets of water with dissolved salts and suspended biological material  ex. Parts of cells from the inside of the mouth



Personnel This respiration-related contamination can be projected large distances from the body Carried by the exhaled air A cough or sneeze can projected the contaminants at high velocity onto critical surface.

Personnel

Eyes can cause problems. The tear ducts regularly produce a saline solution to wet the eyes.

When we blink, small droplets are splashed out of the eyes.



Personnel

 Unfortunately, we occasionally make the human contamination factor worse by adding to the materials which may be shed from the surface of the skin.

Personnel

 Example: Most cosmetics have a powder base which is mixed with an appropriate liquid carrier for ease of application. Once this carrier evaporates, we are left with a semi-dry material which will either flakes off by itself or be carries on skin cell particles.



Personnel Mascara and other eye cosmetics contain large amounts of fine carbon particles which can be shed with time. The colorants in cosmetics are a veritable periodic table with such elements as zinc and iron regularly appearing.



Personnel Men also wear powder-based deodorants and antiperspirants, the latter containing aluminum compounds. The use of aftershave is also problematic, as the alcohol content will dissolve the natural skin oils and thereby allow easier shedding.

 Personnel

◦ A well designed, maintained and operated aseptic process minimizes personnel intervention. ◦ As operator activities increase in an aseptic processing operation, the risk to finished product sterility also increase.

Personnel

◦To ensure maintenance of product sterility, it is critical for operators involved in aseptic activities to use aseptic techniques at all times.