Aseptic Area - Microbiology 1st

Easy to Pharmacy & learnt.
III Sem
B.Pharm - 2nd Year

1. ASEPTIC AREA
It is defined as the area which is free from microbes.
DESIGNING OF ASEPTIC AREA
There are major key points to keep in mind before designing an aseptic area in a manufacturing area
• Adequate space for orderly placement and materials to prevent mix-ups and contamination
• Designing the adequate flow of material and persons to prevent contamination
• Easily cleaned surface and walls e.g. walls should be curved and doors should have no handles
• Adequate and separate areas for various activities (testing, manufacturing)
• HEPA filtered air in manufacturing areas
• Proper flow of manufacturing so as to avoid mix ups

DIFFERENT SOURCES OF CONTAMINATION IN ASEPTIC
AREA AND ITS PREVENTION

There are many sources of contamination which could be due to internal factors, external and manual
1. Internal factors (careless washing of equipment, not following proper dress code, improper
management)
2. External factors (not wearing shoe mask and hair mask etc.)
3. Manual factors (careless handling of the equipment’s and operations like regulating internal
manufacturing environment like temperature, humidity etc.)
Mostly contamination takes place due to carelessness towards handling the equipment’s which could be
avoided by taking certain safety measures.

2. STUDY OF SOURCES OF CONTAMINATION AND
PREVENTION
Prevention of contamination could be achieved if these areas are handled in a
proper way-
1. Personal
2. Buildings and facility
3. Equipment’s
4. Materials
5. Manufacturing process
6. HVAC systems

PERSONAL
Personnel who are supervising or performing drug manufacturing or control can be a potential source of
microbial contamination and a vector for other contamination
• Lack of training
• Inadequate personal cleaning
• Malpractices like eating food, drinking beverages or using tobacco in the storage and processing
area
• Access of unauthorized personnel into production, storage and product control area

BUILDINGS AND FACILITY
Main reasons of contaminations due to improper buildings and facility
• Inadequate filth and pest control
• Rough floors, walls and ceilings
• Lack of air filtration systems
• Improper lighting and ventilation
• Poorly located vents, ledges and drains
• Inadequate washing, cleaning, toilet and locker facility, cleaning of facilities, equipment and
utensils and personal cleanliness

3. EQUIPMENT
Main reasons for contamination from the equipment-
• Improper cleaning and sanitization
• Design preventing proper cleaning and maintenance
• Improper calibration and irregular service
• Deliberate use of defective equipment
• Inappropriate design, size, material leading to corrosion and accumulation of static material and
adulteration with lubrication, coolants, dirt and sanitizing agents


MATERIALS
The raw materials used for production can be a potential source of contamination.
Source of contamination through raw materials
• Storage and handling mistakes causing mix-ups or selection errors
• Contamination with microorganisms or other chemicals
• Improper sampling and testing
• Use of adulterated materials
• Degradation from exposure to excessive environmental conditions such as heat, cold, sunlight,
moisture
• Improper labeling
MANUFACTURING PROCESS
Following are the reasons which causes contamination through manufacturing processes
• Lack of facilities required to manufacture the products
• Use of open manufacturing systems, exposing the product to the immediate room environment
• Lack of cleaning of equipment’s employed in manufacturing.

4. HVAC SYSTEMS
Poor HVAC systems cab be a potential source of microbial growth
• Accumulation of organic material in or near HVAC air intakes
• Ineffective filtration of the supply air
• Insufficient magnitude of pressure differentials causing flow of reversal
• Inappropriate ventilation
Non-directional airflow within production.
MICROBIAL ASSAY
Microbial assay is a sort of bioassays designed to analyze the compounds or substances that have
impact on micro-organisms. They help to estimate concentration and efficiency of antibiotics
Microbial assay is applicable only to the B vitamins. The rate of growth of species of microorganisms
that requires a vitamin is measured in growth media that contain various known quantities of a
foodstuff preparation containing unknown number of vitamins is compared with that obtained from a
known quantity of the pure vitamin
MICROBIAL ASSAY OF ANTIBIOTICS
(OXYTETRACYCLINE)

• Test Organism – Bacillus pumilus
• Culture media is prepared with a pH. of 6.5-6.6 sterile phosphate buffer
• Appropriate conc. Of oxytetracycline is added
• Incubation temperature – 37-39⁰C
• The precision of the assay is such that the fiducial limits of error of the estimated potency (P=
0.95) are not less than 95% and not more than 105% of the estimated potency.

5. AMINO ACIDS
20% of the human body is made up of protein. Protein plays a crucial role in th4e growth of human
body.
Amino acids are basically the building blocks of protein and proteins are the building blocks of human
body
There are 20 different amino acids in nature. All amino acids include 5 basic parts
• A central carbon atom
• A hydrogen atom
• An amino group- consist of nitrogen atom and two hydrogen atoms
• An R- group or side chain- consisting of varying atoms