INTRODUCTION

The sterilization process is the process to reduce or eliminate the contaminant

towards the intended product. Sterilization also known as aseptic technique is a set of
routine measures that are taken to prevent cultures, sterile media stocks, and other
solutions from being contaminated by unwanted microorganisms (Ammirati, 2005). The
sterilization process is significant in medical and laboratory field where the contaminant
may harm the life or contaminate the products. In medical fields, there are cases known
as sepsis, a condition of human body where the outside bacteria produce out the toxins
as their metabolite thus lead to blood poisoning. This case may be prevented with the
proper technique of sterilization.

The contaminants may consist of the 5 categories which the bacteria, yeast,
viruses, fungi and insects. The bacteria are a most frequent contaminant toward the
culture and their interior tissues made them to susceptible to certain aseptic technique
towards a certain extent. The contamination can be categorized in 3 main type which is
initial contamination, latent contamination and introduced contamination. The initial
contamination is due to the inadequate sterilization or just very dirty material. The initial
contamination can be observed in few days where the bacteria will create a turbidity in
the medium where the fungi will look furry in solid medium and a fur ball in liquid medium.
The latent contamination is a contamination that can be observer after a long time of
cultures and it is usually the bacterial contamination. Apparently the bacteria are present
endogenously in the initial plant material and are not obviously pathogenic in situ. It is
dangerous it can be transferred between the cultures (AFBI, 2011). The introduced
contamination then caused by the improper handling of cultures or the non-sterilize
compound. It can be prevented through the proper aseptic technique.

One of leading sterilization process in for the sterilize preparation is the autoclave.
Autoclaving is the method most often used for sterilizing heat-resistant items and our
usual method for sterilizing items. In order to be sterilized, the item must be held at 121°C,
15 psi, for at least 15 minutes. It is important that items reach this temperature before
timing begins. Empty vessels, beakers, graduated cylinders, etc., should be closed with
a cap or aluminum foil. Tools should also be wrapped in foil or paper or put in a covered
sterilization tray. It also a crucial point to let the steaming to penetrate inside the glassware
to increase the sterilization rate. (AFBI, 2011). Based on the “Environmental Health &
Safety Fact Sheet: Autoclaved Waste", types of waste that should not be autoclaved
include: cancer drugs, toxic chemicals, radioisotopes, volatile chemicals or any other
harmful material that can be vaporized and disseminated with heat. In general, do not
autoclave flammable, reactive, corrosive, toxic or radioactive materials. There are several
safety precautions that need to obey when handling the autoclave machine. Autoclaving
produces steam, which can cause burns and requires a lot of caution in handling. The
most important moment is opening the machine after completion of the cycle. Wear the
standard PPE when operating the autoclave. Check that the chamber pressure is zero
before attempting to open the door. Stand behind the door when opening and use it as a
shield. Slowly open a crack and when removing liquids from the autoclave, point the
opening of the container away from yourself and other people because the liquid may
suddenly boil and spray out the opening (Ammirati, 2005).

A laminar flow unit (or hood) is a sophisticated appliance that can further help pre-
vent contamination of reagents and biological cultures (Ammirati, 2005). A horizontal
laminar flow unit is designed to remove particles from the air. It also keeps room air from
entering the work area and both suspends and removes airborne contaminants
introduced into the work area by personnel. Room air is pulled into the unit and pushed
through a HEPA (High Energy Particle Air) filter with a uniform velocity of 90 ft/min across
the work surface. The first HEPA filters was created on 1940s, when the USA Atomic
Energy Commission under Manhattan Project decide to developed an efficient and
effective way to filter the radioactive particulate contaminants (Ammirati, 2005). The air is
filtered by a HEPA (high efficiency particulate air) filter so nothing larger than 0.3
micrometer, which no bacterial and fungal spores can pass through. The positive
pressure of the air flow from the unit also discourages any fungal spores or bacteria from
entering. Depending on the design of the hood, the filters are located at the back or in the
top of the box (AFBI, 2011).
 Figure 1: Transfer hood air flow (AFBI,2011)

Many laminar flow hoods are equipped with ultraviolet (UV) lights, which
can help lessen contamination by inducing DNA damage to potential contaminants. Such
light is also harmful to laboratory workers. The safety precaution when operating UV light
is never look directly at a UV light, as that can cause eye damage. The UV light can also
cause skin burns (sun burns) to anyone in the room. Consequently, UV lights should
always be turned off whenever the room is occupied (Ammirati, 2005). Laminar flow
hoods are essential components of many biosafety level (BSL)-2 laboratories, where they
help prevent spread of viruses and some bacteria.
OBJECTIVES

1. To identify the components of sterilization equipment such as laminar air flow

cabinet and autoclave machine
2. To study the principle and operation for both the laminar flow cabinet and
autoclave.

REFERENCES

AFBI. (2011). 9. Contaminants. State of the Seas Report, 1–6.

Ammirati, C. T. (2005). Aseptic Technique. Surgery of the Skin, (November), 25–37.
https://doi.org/10.1016/B978-0-323-02752-6.50007-9
ENVIRONMENTAL HEALTH & SAFETY FACT SHEET: AUTOCLAVED WASTE. (n.d.).
Biological Safety Services, 9244.