STERILE PRODUCTS AND MICROBIOLOGY

Mahesh Dulange
M.Sc. (Microbiology), LSSGB/LSSBB
 STERILE PRODUCTS AND MICROBIOLOGY

Why are Some Products Required to be Injectable…?

Choice is based on:
 Therapeutic need
 Speed of response
 Specificity of response needed
 Concerns on Absorption, Distribution, Metabolism andExcretion
 Stability of formulation including interactions withcontainer and excipients
Sterile products :
 Bypass the skin, an effective barrier to infection, and come into direct contact with
internal tissues and fluids where infection can easily arise. Sterility of these
preparations is an absolute requirement
 onset of action is usually very rapid and very difficult to reverse
 Patients receiving these products can be very ill or immunocompromised. Many
of these products are truly ‘life saving’
 defective product can kill a patient!
Sterile products come in many different shapes and sizes including:
 Large and Small Volume Parenterals (LVPs and SVPs) filled into ‘traditional’
plastic infusion bags or glass vials and ampoules
 Eye ointments
 Pre-filled syringes

How are Sterile Products CommonlyAdministered?

 Subcutaneous-most common, just underneath the dermis and epidermis
 Intravenous-directly into venous system (blood), highest risk
 Intramuscular-directly into muscle, more painful
 Intrathecal-into the spine
 Intracardial-into heart
 Intracerebral-into brain
 Ophthalmic formulations- into eyes
 Other : Topical formulations (Wound cream)

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 STERILE PRODUCTS AND MICROBIOLOGY

The ‘quality requirements’ for the sterile products are most demanding and stringent in
the pharmaceutical industry. Mainly those are:
 Sterility :Free of all living forms
 Endotoxins: Some types of microorganisms, mainly the Gram negative water
borne species, can cause a temperature rise in patients due to endotoxins. This
‘pyrogenic’ response can be potentially fatal for patients already very ill. Pyrogens
(or Endotoxins) originate from the outer cell wall of these microorganisms when
they are both alive/dead. They are also very difficult to remove.
 Low in B that may block very small capillaries
 Chemical and Biological purity
 Container integrity to prevent ingress of microorganisms. The container must
remain intact throughout its shelf life
 Stable
 Correctly labelled. Although often ignored, a mislabelled product can be just as
life threatening as a contaminated one.
Microbiology is that part of biology dealing with the smallest of living things, the
microorganisms. Microorganisms can be broadly grouped into:
 Bacteria
 Fungi

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 STERILE PRODUCTS AND MICROBIOLOGY

 Algae
 Protozoa
 Viruses
Microorganisms are widespread within the environment in very high numbers, often
surviving in very harsh environmental conditions with minimum nutrition. Two rich
sources of microorganisms are the soil and the human body. For example: Each square
centimetre of skin contains approximately 1 million bacteria, with saliva containing ~
109 microorganisms per millilitre In the pharmaceutical industry the microorganisms of
particular interest are:
Bacteria-Gram negative, spore formers (Disinfectant resistant)
Fungi-spore formers (Disinfectant resistant) hard to remove
 Bacteria:
Bacteria multiply by binary fission, which means that one cell divides into two, which in
turn becomes four, then eight, then sixteen…. and so on. Actively growing bacteria have
a generation time of 20-30 minutes, which means that one cell can multiply to millions
within hours. One cell of the bacterium E.coli could theoretically, if sufficient food were
available, produce a mass of bacteria greater than the mass of the Earth ( 5.972 X 10 24 kg)
in just 3 days!!
Bacterial Cell Wall – Characteristics
Gram positive bacteria 20‐80nm thick
Gram negative bacteria 5‐10nm thick
Additional outer membrane (slime/capsule)
Outer membrane characteristics
Rich in lipopolysaccharide and lipoprotein
Key mechanism of attachment(biofilm formation )
Cell wall provides structural integrity
Cell wall characteristics used in identification –
Gram positive and gram negative based on gram reaction
Gram positive organisms are a very common environmental contaminant
The presence of Gram negative organisms indicates that water is present somewhere in
the environment
Gram negative organisms are source of endotoxins.
When Gram positive bacilli are found there is a theoretical risk of resistant endospores.

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 STERILE PRODUCTS AND MICROBIOLOGY

Bacteria are the most common source of contamination in the cleanroom , where the
sterile products area manufactured and filled.
 Fungi: Fungi can also contaminate pharmaceutical products and the manufacturing
environment. Fungi occur in two forms:
 Moulds such as aspergillus
 Yeasts such as Candida
Characteristics of Moulds
Produce spores for reproduction, dissemination and protection. Unlike spore forming
bacteria, where only one spore is produced per cell, moulds can produce many thousands
of spores. These spores are not as resistant as bacterial endospores
Nutritional requirements are generally simpler than those of bacteria. They can also grow
in the absence of significant amounts of water
Moulds require a longer incubation period for growth and specialised methods of
identification and staining
Moulds are widespread in the environment, present in soil and decaying vegetation. They
can also be found in cardboard shippers, exposed plaster and damaged pipework lagging
They are second only to bacteria as the most common source of contamination in the clean
room. Under some circumstances, they can become a serious threat.
Characteristics of Yeasts
Appear as large Gram positive spheres/ovoids following staining
A few yeasts are capable of producing moderately resistant spores as a means of
reproducing. More commonly, however, they reproduce by producing less resistant ‘buds’
Generally require high levels of nutrition and moisture. Although rarely a contaminant in
a clean room, they can cause spoilage of pharmaceutical products that contain high levels
of water and carbohydrates.
 Nutritional Requirements – Variable
Microorganisms’ requirement for food and nutrition are extremely variable:
Pseudomonas species only require 1ppb of carbon to survive… they easily grow in
purified water
Bacillus can survive on paper and cardboard
Moulds can readily survive on wood and in plaster
Pseudomonas = Water source
Bacillus = Environmental source

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Moulds = Environmental source

Staphylococcus = People source
Although they are on us, inside us and around us in very high numbers they are difficult
to capture and isolate
… Just because they aren’t in the sample does not mean they will not be.
Hence , avoiding the entry of microorganisms by means od disinfection, sterilization
and aseptic practices in the clen room area are the only way to protect the patient an
the product.
 Recent recalls :
 24 FEB 2023- global pharma -Artificail eye ointment due to possible microbial
contamination
 03 March 2023-pharmediaca USA LLC- 15% MSM drops- non sterility
 03 March 2023- Apotex- Brimonidine Tartrate opth. Solution -Lack of sterility

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