PHCT 401 Aseptic Processes & Techniques · 2017-01-12 · PHCT 401 Aseptic Processes & Techniques...

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PHCT 401 Aseptic Processes & Techniques Principles of Aseptic Techniques Aseptic Processing Sterility Testing Laminar flow air cleaning Quality Control Tests Personnel

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Transcript of PHCT 401 Aseptic Processes & Techniques · 2017-01-12 · PHCT 401 Aseptic Processes & Techniques...

  • PHCT 401

    Aseptic Processes & Techniques

    Principles of Aseptic Techniques

    Aseptic Processing

    Sterility Testing

    Laminar flow air cleaning

    Quality Control Tests


  • Principles of Aseptic Techniques


    • Certain pharmaceutical products must be sterile

    • Parenteral preparation o Injections

    o Intravenous infusions

    o Non-injectable sterile fluids Urological (bladder) irrigation solutions

    Peritoneal dialysis & Haemodialysis solutions

    • Ophthalmic preparations o Eye drops/Oint

    o Eye lotions

    o Contact lens solution

  • • Dressings

    • Implants

    • Two categories of sterile products

    Terminally Sterilized (TS) – can be sterilized in

    their final containers

    Non-Terminally Sterilized (NTS) – cannot / will

    not be sterilized in their final containers.


    ‘Starting with sterile materials and equipments, it is possible to produce a sterile product if strict

    precautions are taken to avoid microbial


  • • Methods used, collectively described as “Aseptic Technique”

    • Aseptic manufacturing procedures available for different types of products.

    • Awareness of aseptic handling procedures to be adopted to minimize risk of product contamination.

    • Those who have cause to open, use or dispense sterile products e.g Hospital Pcy

    • Manufacturing under conditions that do not permit entry of contaminating µorganisms

    Aseptic Manufacturing Processing

  • Asepsis

    “A state of control attained by using an

    aseptic work area and performing activities

    in a manner that precludes microbiological

    contamination of the exposed sterile


    Aseptic Processing

    “the processing of drug components (drug,

    containers, excipients etc.) in a manner that

    precludes microbiological contamination of

    the final sealed product”

  • Aseptic processing:

    “A method of producing sterile products in which sterile bulk product or sterile raw materials are compounded and filled into sterile containers in a controlled environment, in which the air supply, materials, equipment and personnel are regulated to control microbial and particulate contamination to acceptable levels”

    • Objective is to maintain the sterility of a product, assembled from sterile components

    • Operating conditions so as to prevent microbial contamination.

    • Combination of various simple techniques and processes.

  • • Spoilage of medicines due to microbial

    contamination is undesirable

    • Various consequences

    Financial loss – main

    Risk of initiating infection (though uncommon)?

    More important in terms of

    oRisk to the patient

    oPossible loss of life

    Financial implications of product-related infections

    Additional treatment costs

    Product recalls

    Possible litigation & damage to reputation

  • Understanding Aseptic processes & techniques

    • Understand various sources of contaminating microorganisms in the production or work area

    Personnel o Skin and respiratory tract flora

    oMicrobial transfer from operators

    Atmosphere - air


    Raw materials

    Packaging materials


    Buildings oWalls & ceilings

    o Floors &drains

    o Doors, windows & fittings

  • • Quality of pharmaceutical products refers to ‘fitness for purpose’

    • Product should not only have the desired therapeutic properties

    • Safe for administration by intended route

    • Sterile products – free from microbes

    • Others like oral preparations, need not be sterile

    Free from indicator pathogens that can be contracted through the oral route

    • Greater attention paid to quality of sterile product

    • Reflects additional Quality assurance required

  • • In demonstrating quality, tests carried out to detect the absence of quality

    • Sterility Test

    o Sampling products at random

    o Testing for presence of microorganisms

    o Samples taken should be representative of the whole population –Sampling Technique

    o Absence of microorganisms will only allow you to estimate the statistical probability that the batch sampled is sterile.

    o All products should therefore be manufactured in a suitable environment

    By procedure that minimizes the possibility of microbial contamination occurring

    Building quality into the product

    At the end of the manufacturing process, tests can be performed as additional measure, since

    Quality is not inspected into the product

  • Application of Aseptic Techniques & Processes

    Techniques designed to prevent accidental contamination of sterile materials & pure cultures from the environment during handling

    • Pharmaceutical manufacturing

    • Foods & cosmetics

    • Pure cultures of microorganisms used in Pharmaceutical biotechnology

    Recombinant human insulin

    Production of immunological products- vaccines

    Production of antibiotics

    Careful handling to prevent accidental contamination of pure stock

  • Aseptic technique involve procedures such as:

    • Use of sterile apparatus & materials at all


    • Use of proper closures & containers

    • Wrapping of apparatus for sterilization

    • Flaming of necks of containers in Bunsen

    burner flame

    • Minimal exposure of materials during handling

    • Spraying & swabbing of benches & work area

    with a suitable disinfectant

    • Washing & scrubbing of hands

  • Keeping the operator’s dirt and germs out of the sterile cleanroom environment and away from sensitive products and processes is the main objective of the sterile cleanroom suit

    Automated Hand-Washing System

    •Use of protective clothing-to keep all clothing

    parts together

  • Carrying out operations within/under special handling

    cabinets equipped with filtered air.

  • Wearing of protective clothing

    • Clothing worn must be made from non-

    shedding materials: terylene a suitable fabric

    • Proper head gears/wears

    • Rubber or plastic gloves

    • Face masks to prevent the release of droplets

    must be worn by operators in the aseptic

    areas for production of especially NTSP

    • Prevents airborne contamination of both

    microbial and particulate matter.

    • Clothing that are close fitting at the neck, wrist

    and ankles more suitable

  • • Clothing should be used once a day

    • Fresh headwear, overshoes and powder-free

    gloves should be provided for each working


    Four Pillars of a robust Aseptic Processing

    • Personnel Training &Monitoring

    • Environmental Monitoring

    • Facility Design HVAC Validation

    • Process Simulation (Media Fills)

  • Sterility Testing

    • Required for all sterile products

    • Not overriding control to cover all short-


    • Adequate attention must be paid to every

    stage in the production process

    Adequate premises

    Sterilizing equipment and process

    Skilled and trained personnel etc

    • Building quality into the product

    • ST is actually the detection of the absence of


  • • Sterility does not detect all known living forms

    • Official tests for sterility detects gross contamination

    with most common organisms (including pathogens)

    • Most probable contaminants limited to bacteria and


    • No test for viruses

    • Difficulty in culturing viruses

  • Design of Sterility Tests

    Critical considerations

    Sampling – method should give the greatest

    probability of picking up an infected unit within the



    Container Unit

    Volume of medium

    Inhibitory substances in the product

    Product should not affect the ability of the medium to

    support growth.

    Added product not > 10% of the volume of medium. B.P

    Details from U.S.P.

  • Media – appropriate to support likely contaminant


    Fungi – modified


    Of preservatives

    Any constituent with AMA activity

    Inactivation aids recovery of contaminants


    • Not inhibitory to growth promoting property of


    • Should not interfere with nutritive quality of the


  • Methods of inactivation

    • Dilution

    • Inactivation by constituents

    • Destruction or inactivation / antagonization by

    addition of sterile inactivators.

    • Physical removal by filtration.

    Other critical consideration

    • Controls

    • Incubation conditions

    • Interpretation of results

  • Controls

    • Two major controls against which to check

    conclusion from the test series.

    • Negative Control

    Sterility of the media used

    Checked by including un-innoculated bottles from

    the same batch as that used in the test

    • Positive Control

    Growth promoting properties of the batch

    Inactivation of inhibitory substances in the product

    Demonstrated by including an exact set with same

    quantities of test substance (sample) in the same

    volumes of the batch of test media

  • Reporting Sterility Testing Results









    AET Sample Thioglycolate No Organism - ve

    ANT Sample Thioglycolate No Organism -ve

    FGT Sample SDM No Organism - ve

    AET PC No Sample Thioglycolate S. aureus + ve

    ANT PC No Sample Thioglycolate Clostridium + ve

    FGT PC No Sample SDM Candida + ve

    AET NC No Sample Thioglycolate No Organism - ve

    ANT NC No Sample Thioglycolate No Organism -ve

    FGT NC No Sample SDM No Organism - ve


    AET = Aerobic Test

    ANT = Anaerobic Test

    FGT = Fungal Test

    PC = Positive Control

    NC = Negative Control

  • Laminar Flow Air Cleaning

    • Providing the environment for aseptic manufacturing

    • Cleanrooms with specific requirements

    • Two major types

    • Differentiated by their method of ventilation.

    Turbulently ventilated cleanrooms (‘nonunidirectional’)

    Unidirectional flow cleanrooms (originally known as

    ‘laminar flow’

    The unidirectional type of cleanroom uses very much more

    air than the turbulently ventilated type,

    Gives a superior cleanliness. w.

  • Non-unidirectional Airflow

    Figure shows a turbulently

    ventilated room receiving clean

    filtered air through air diffuses in

    the ceiling. This air mixes with the

    room air and removes airborne

    contamination through air extracts

    at the bottom of the walls. The air

    changes are normally equal to, or

    greater than, 20 per hour, this

    being much greater than that used

    in ordinary rooms, such as in

    offices. In this style of cleanroom,

    the contamination generated by

    people and machinery is mixed

    and diluted with the supply air and

    then removed.

  • Unidirectional Airflow The horizontal flow cleanroom uses the same filtration airflow technique as the downflow, except the air flows across the room from the supply wall to the return wall. One major limitation of the horizontal flow design is that the downstream contamination in the direction of airflow increases.

  • Steps in Aseptic Room Air Filter

    • Air intake

    • Pre-filtration

    • Temperature regulation

    • Humidification

    • HEPA filtration

    HEPA - High Efficiency Particulate Air filters

    Can remove 99.97% of airborne

  • Air intake

    • From environment – high up

    • Away from polluted area of town & chimneys


    • Through a pre-filter

    • Set of coarse filters next to the air intake

    • Removes large particles (≥5µm)

    Temperature regulating/adjustment

    • Regulates air temperature to ambient

    • If too cold, heats up

  • Humidification

    • Prepare air of required humidity

    • Incoming air passed through system of fine atomized

    spray of de-mineralized water

    HEPA filtration

    • Final filtration step

    • Air forced through high efficiency air filters

    • Removes 99.97% of airborne particles of 0.3µm size

    • Particles 0.3µm size most difficult to filter

    • Considered MPPS (Most Penetrating Particle Size)

  • Airflow Control

    • Vertical laminar flow

  • Personnel

    • High standards of personal hygiene

    • Most common source of contaminants in

    sterile drug products.

    • Large numbers of microbes

    • Healthy population known as reservoir of

    many microbes.(pathogens)

    • Shed numerous ‘viable’ & ‘non-viable’

    particles into environment

    • Free from communicable diseases & open

    lesions on exposed body surfaces.

  • • Organisms carried on (larger) skin particles.

    • Adherence to high standards of cleanliness

    ensured by

    Adequate hand-washing facilities

    Protective garments – head gears,


  • Appropriate training of all staff before being

    permitted to enter the Aseptic Manufacturing


    Fundamental Training Topics

    Principles of cGMP

    Practice & theory of assigned task

    Aseptic technique


    Gowning proficiency – training, observation.

    Specific SOPs covering aseptic

    manufacturing area operations.

  • Laminar Flow Air Cleaning