UNIVERSITY OF GONDAR

INSTITUTE OF TECHNOLOGY
DEPARTMENT OF BIOMEDICAL ENGINEERING
TITLE: STERILIZATION AND REFRIGRATION SYSTEM

GROUP MEMBERS
NAME ID-NO

1. GETAW ZEWUGE........................................GUR/00774/10

2. ENDALE ZEBENE......................................GUR/00761/10

3. TAREKEGN TILA......................................GUR/00481/10

4. KEFYALEW GETAHUN..........................GUR /00858/10

5. YARED MELESE........................................GUR /01158/10

6. ABEBE BELETE.........................................GUR/01012/10

SUBMMITED TO MATIWOS.T
SUBMMISITION DATE 18/08/2013
Contents
STERILIZATION.............................................................................................................................................1
Classification of Materials to be sterilized...................................................................................................1
Producer of sterilization..............................................................................................................................2
Methods of sterilization..............................................................................................................................4
Heat sterilization.........................................................................................................................................4
Autoclave (steam sterilization)................................................................................................................5
Dry-heat sterilization...............................................................................................................................7
Radiation sterilization..................................................................................................................................9
Chemical sterilization................................................................................................................................11
Refrigerating system in hospital................................................................................................................14
Pharmacy refrigerating system..............................................................................................................16
Lab Refrigeration...................................................................................................................................16
Refrigerator of blood in bank................................................................................................................17
REFERENCE................................................................................................................................................18
STERILIZATION

Sterilization is the complete destruction or removal of all microorganisms (including spore-

forming and non Spore forming bacteria, viruses, fungi, and protozoa) that could contaminate
pharmaceuticals or other materials and thereby constitute a health hazard. Sterilization is an
operation that eliminates or kills micro-organisms borne on inert media. The result of this
operation is a “sterile state” in which the survival of a micro-organism is highly unlikely. A
suitable limit for this probability has been defined (based on studies by NASAand the WHO) as
being 10-6(one per million), meaning the probability of one in a million of finding a viable micro-
organism on a sterilized item.

Since the achievement of the absolute state of sterility cannot be demonstrated, the sterility of a
pharmaceutical preparation can be defined only in terms of probability. The efficacy of any
sterilization process will depend on the nature of the product, the extent and type of any
contamination, and the conditions under which the final product has been prepared. The
requirements for Good Manufacturing Practice should be observed throughout all stages of
manufacture and sterilization. ( Methods of sterilization , Ninth Edition, 2019)

Furthermore, different micro-organisms have varying resistance to heat, and some will require
much longer exposure to heat than others. This means that the various micro-organisms found on
the surface of an instrument will not be destroyed at the same time, hence why the stated
sterilization time must be obeyed.

Classification of Materials to be sterilized

Critical Items

Critical items confer a high risk for infection if they are contaminated with any microorganism.
Thus, objects that enter sterile tissue or the vascular system must be sterile because any microbial
contamination could transmit disease. This category includes surgical instruments, cardiac and
urinary catheters, implants, and ultrasound probes used in sterile body cavities etc.

Semi-critical Items

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Semi-critical items contact mucous membranes or non-intact skin. This category includes
respiratory therapy and anesthesia equipment, some endoscopes, laryngoscope blades,
esophageal manometer probes, cystoscopes, anorectal manometry catheters, and diaphragm
fitting rings etc.

Noncritical Items

Noncritical items are those that come in contact with intact skin but not mucous membranes.
Intact skin acts as an effective barrier to most microorganisms; therefore, the sterility of items
coming in contact with intact skin is “not critical.”

They can be Non-critical patient care items: bedpans, blood pressure cuffs, crutches and
Computers Non-critical environmental surfaces. (William A. Rutala, 2016 Aug 9)

Producer of sterilization

A better perceptive of sterilization has shown us that sterilizing an instrument does not mean
merely autoclaving or steam sterilizing it. Somewhat, sterilization consists all the treatment
actions that medical devices require after use to make sure they pose no risk of infecting the next
patient. The sterilization process includes the following steps:

Pre-disinfection.

Detergence-disinfection.

Conditioning

Sterilization by autoclaving or any other method.

Load release after verification

Storage and availability for use

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Pre-disinfection

Pre-disinfection consists of soaking the equipment in an effective, fast-acting detergent-

disinfectant for at least 15 minutes. The detergent-disinfectant must be non-corrosive to the
equipment and have a low toxicity to staff. Pre-disinfection can achieve reductions of 10 -3(one
per thousand) in the number of germs on the surface of an item, and is the only step in the
sterilization process that may be performed outside of the sterilization department. Pre-
disinfection in a ward requires a location where the procedure can be performed in proper
conditions (e.g. with space, a water supply, a water drainage system, and adequate staff and
equipment).

Detergence-disinfection

This second disinfection step is important and achieves further reductions of 10 -5 in the number
of germs on the surface of an item requiring sterilization. Detergency-disinfection combines
mechanical brushing with the chemical action of a detergent-disinfectant.

Conditioning

The material used for conditioning must: allow steam to penetrate during sterilization and Keep
instruments sterile after autoclaving until they are used on a patient; the material must thus be
impermeable to micro-organisms.
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Instruments are wrapped in sheets of crepe sterilization paper or other non-woven paper.
Sterilization sheets are single-use, and must never be reused. Exposure to steam and to
differences in pressure during autoclaving alters the physical properties of sterilization sheets.

Sterilization in an autoclave or another technique

After such producers we have to use any appropriate sterilization techniques physical method
like heat sterilization (steam heat or dry heat) radiation sterilization or chemical sterilization in
order to make the instrument or the surface sterile.

Load release

Sterilization unit technicians may not release the load. Authorize its use until they have carried
out specific checks. Dryness and integrity

When removing a load from an autoclave or other technicians must ensure that the load is dry
and the conditioning sheets are not torn.

Sterilization quality

It is impossible to define a target sterility value for medical devices at the end of the sterilization
procedure, before their use. Reliable methods are therefore required to determine whether a
medical device is sterile.

Storage and availability

Storage conditions are important for maintaining sterility. The sterilized packets must be
protected from dust and any possible damage. (Pascale Choquenet, 2017)

Methods of sterilization

 Heat sterilization
 Radiation sterilization
 Chemical sterilization

Heat sterilization

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Heat sterilization is the mainly used and reliable method of sterilization, concerning destruction
of enzymes and other essential cell constituents of microorganisms. The process is more
effective in hydrated state where under conditions of high humidity, hydrolysis and denaturation
occur, thus lower heat input is required. Under dry state, oxidative changes take place, and
higher heat input is required. Heat sterilization techniques include:

Autoclave (steam sterilization)

Introduction of microorganisms to saturated steam under pressure in an autoclave achieve their

damage by the irreversible denaturation of enzymes and structural proteins of microbes. The
temperature at which denaturation occurs changes inversely with the quantity of water present.
Sterilization in saturated steam thus requires accurate control of time, temperature, and pressure.
As disarticulation of the air by steam is unlikely to be readily achieved, the air should be
evacuated from the autoclave before admission of steam. This method should be used whenever
possible for aqueous preparations and for surgical dressings and medical devices.

The temperature should be used to manage and watch the process; the pressure is mainly used to
acquire the necessary steam temperature. (Methods of sterilization, 2019)

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Optional conditions, with varies combinations of time, pressure and temperature, are given
below.
Take that 1 atm = 101 325 Pa

Temperature (°C) Approximate Minimum sterilization time

corresponding pressure (min)

(kPa)
126-129 250 (~2.5 atm) 10
134-138 300 (~3.0 atm) 5

The Steam Sterilization Cycle

A steam sterilization cycle consists of three basic phases:

Heating (come-up) phase - steam enters the sterilizer jacket and air is removed from the
sterilizer chamber, either by gravity displacement or mechanically (Prevacuum). Objective:
Remove air and replace with steam.

Sterilization (exposure) phase - load is exposed to steam at a set temperature (Measured and
controlled by a temperature sensor in the drain line) for a set time. Objective: To sterilize the
product.

Cool-down (drying) phase - sterilizer chamber is exhausted to atmospheric pressure followed

by circulating air through the chamber or by drawing a deep vacuum. Jacket heat is maintained
during the drying phase. Objective: To return the product to atmospheric pressure and to help dry
the product (prevacuum and gravity cycles).

Critical process Autoclave parameters:

 TIME

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  TEMPERATURE
 SATURATED STEAM. (Butler, August 2009)
• Advantages
 Non-toxic
 Cycle easy to control and monitor
 Inexpensive
 Quick microbe kill times
 Least affected by various soils
 Rapid cycle time
 Penetrates medical packing, hollow tubing
• Disadvantages
 Not for materials that are sensitive to heat or moisture
 Potential for burns
 Steam generator system is needed (Butler, August 2009)

Dry-heat sterilization

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Dry-heat process is considered to be oxidation of cell constituents. Dry-heat sterilization needs a
higher temperature than steam sterilization and a longer experience of time. The method is,
therefore, more suitable for heat-stable (heat resistant), non-aqueous (insoluble) materials that do
not sterilized by steam because of its deleterious effects or failure to penetrate. Such materials
contain glassware, powders, oils, and some oil-based injectables. Preparations to be sterilized by
dry heat are filled in units that are either sealed or temporarily closed for sterilization. The entire
content of each container is maintained in the oven for the time and at the temperature given in
the table below. Other conditions may be necessary for different preparations to ensure the
effective elimination of all undesirable microorganisms.

Temperature Minimum sterilization time

(°C) (min)

160 180
170 60
180 30

The oven should normally be equipped with a forced air system to ensure even distribution of
heat throughout all the materials processed. This should be controlled by monitoring the
temperature. Containers that have been temporarily closed during the sterilization procedure are
sealed after sterilization using aseptic techniques to prevent microbial recontamination. (Methods
of sterilization, 2019)

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Radiation sterilization

Radiation is currently used for sterilization and decontamination in the medical supplies (surgical
supplies, vaccines, and drugs) and food industries. Two types of radiations are ionizing and non-
ionizing are efficiently used.

Ionizing radiation

Ionizing radiation is an excellent agent for sterilization/disinfection; it kills organisms without

increasing the temperature; so aptly called cold sterilization. It destroys bacterial endo sore and
vegetative cells, both eukaryotic and prokaryotic; but not always effective against viruses.

Mechanism of ionizing radiation

When ionizing radiation collides with particles they produce electrons and other such as
hydroxyl radicals and hydride radicals Each of these reactive molecules is capable of degrading
and altering biopolymers such as DNA and protein Breakage of DNA and degradation of
enzymes lead to the death of the irradiated cells.

Several sources of ionizing radiation are available, including X-ray machines, cathode ray tubes
(electron-beam radiation), and radioactive nuclides (sources of gamma/x-rays).

Cathode Rays (Electron-Beam Radiation)

Cathode rays or electron-beams can sterilize materials at room temperature with brief exposure.
They have limited penetrating power and are used for the sterilization of surgical supplies, drugs,
and other materials.

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Gamma sterilization

Gamma radiation is used in the sterilization of;

Disposables such as plastic syringes, infusion sets, catgut sutures, catheters, gloves, and adhesive
dressings before use.

Bone, tissue grafts, antibiotics, and hormones.

Irradiation of food (permitted in some countries).

Advantages of Ionizing Radiations sterilization

High penetrating power: products can be processed in their fully sealed, final packaging thus
limiting the risk of contamination following sterilization.

Rapidity of action: saves and efforts.

Temperature is not raised: compatible with temperature-sensitive materials, such as

pharmaceuticals and biological samples.

Flexibility: can sterilize products of any phase (gaseous, liquid, or solid materials), density, size,
or thickness.

Disadvantages

Capital costs are high and specialized facilities are often needed e.g. for gamma irradiation.

Use of gamma radiation requires handling and disposal of radioactive material.

Not compatible with all materials and can cause breakdown of the packaging material and/or
product

Non-ionizing Radiation

Non-ionizing radiations are quite lethal but do not penetrate glass, dirt, films, water; hence their
use is restricted for disinfection of clean surfaces in operation theaters, laminar flow hoods as
well as water treatment. The recommended dose is 250-300 nm wavelengths, given for 30
minute.

Infra-Red Radiation

Infra-red rays are low energy type electromagnetic rays, having wavelengths longer than those of
visible light. They kill microorganisms by oxidation of molecules as a result of heat generated.
Infra-red rays are used for the rapid mass sterilization of syringes and catheters.

Ultraviolet Light (UV) Sterilization

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Sunlight is partly composed of UV light but shorter wavelengths of light are filtered out by the
ozone layer. There are three types of UV radiation; UVA, UVB, and UVC, classified according
to their wavelength. Short-wavelength UVC is the most damaging type of UV radiation.

Mechanisms of UV Sterilization

Many cellular materials including nucleic-acids absorb ultraviolet light. It causes bonding of two
adjacent pyrimidines i.e., the formation of pyrimidines dimer, resulting in the inhibition of DNA
replication. This leads to mutation and death of exposed organisms.

Uses of UV Sterilization

UV lights are useful for sterilization surfaces, air, and water that do not absorb the UV rays.
Certain types of UV lights can kill influenza virus. Ultraviolet radiation is used for disinfecting
enclosed areas such as bacterial laboratory, nurseries, inoculation hood, laminar flow, and
operation theaters. For illustration, laboratory biological cabinets all come equipped with a
“germicidal” UV light to clean up the surface after use.

Disadvantages

Damages skin and eyes: Conventional UV light can penetrate and damage skin and also the
reason for cataracts.

Does not penetrate paper, glass, and cloth. (Tankeshwar, April 24, 2020)

Chemical sterilization

Chemical Sterilization is the process of elimination of microorganisms by the use of chemical

bactericidal agents. Even if physical methods like heat sterilization are more proper for effective
sterilization, sometimes it may not be suitable to use for heat-sensitive materials like plastics,
fiber optics, and biological specimens.

For such situation, chemical either in liquid or gaseous state can be used for sterilization.
However, it is necessary to ensure that the materials used for sterilization are well-matched with
the chemical being used. We have to accept safety considerations in the workplace safety during
the use of chemical agents.
The chemical method of sterilization can be classified as liquid and gaseous sterilization.
Gaseous sterilization

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 Gaseous sterilization is the process of exposing equipment or devices to different gases in a
closed heated or pressurized chamber (container).
 Gaseous sterilization is a more effective technique because gases can pass through a tiny
orifice and provide more effective results.
 Besides, gases are commonly used along with heat treatment which also facilitates the
functioning of the gases.
 However, there is an issue of release of some toxic gases during the process which needs to
be removed regularly from the system.
The following are some of the common gases used for gaseous sterilization
I. Ethylene oxide
 Ethylene oxide (EO) gas is a common gas used for chemical treatment applied to sterilize,
pasteurize, or disinfect different types of equipment and surfaces because of its wide range
of compatibility with different materials.
 EO treatment often replaces other sterilization techniques like heat, radiation, and even
chemicals in cases where the objects are sensitive to these techniques.
 This method is a general method used for almost 70% of all sterilizations and
approximately 50% for disposable medical devices.
 Ethylene oxide kills all known microorganisms, such as bacteria (including spores),
viruses, and fungi (including yeasts and molds), and is compatible with almost all materials
even when constantly applied.
II. Formaldehyde
 Formaldehyde is another important highly reactive gas which is used for sterilization. This
gas is obtained by heating formalin to a temperature of 70-80°C.
 It possesses broad-spectrum biocide activity and has found application in the sterilization
of reusable surgical instruments, specific medical, diagnostic and electrical equipment, and
the surface sterilization of powders.
 Formaldehyde doesn’t have the same penetrating power of ethylene oxide but works on the
same principle of modification of protein and nucleic acid.
 As a result of the low penetrating power, its use is often limited to paper and cotton fabrics.

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 Formaldehyde can generally be detected by smell at concentrations lower than those
permitted in the atmosphere and thus can be detected during leakage or other such
accidents.
III. Nitrogen dioxide
 Nitrogen dioxide is a rapid and effective sterility that can be used for the removal of
common bacteria, fungi, and even spores.
 NO2 has a low boiling point (20°C) which allows a high vapor pressure at standard
temperature.
 An advantage of this gas is that no condensation of the gas occurs on the surface of the
devices because of the low level of gas used and the high vapor pressure. This avoids the
need for direct aeration after the process of sterilization.
IV. Ozone
 Ozone is extremely reactive industrial gas that is usually used to sterilize air and water and
as a disinfectant for surfaces.
 Ozone is a potent oxidizing property that is capable of destroying a wide range of
organisms including prions, without the use of hazardous chemicals as ozone is usually
generated from medical-grade oxygen.
 Similarly, the high reactivity of ozone allows the removal of waste ozone by converting the
ozone into oxygen by passing it through a simple catalyst.
 However, because ozone is an unbalanced and reactive gas, it has to be produced on-site,
which limits the use of ozone in different settings.
 It is also very dangerous and thus only be used at a concentration of 5ppm, which is 160
times less than that of ethylene oxide.
Liquid sterilization
 Liquid sterilization is the process of sterilization which involves the submerge of
equipment in the liquid sterile to kill all viable microorganisms and their spores.
 Although liquid sterilization is not as effective as gaseous sterilization, it is suitable in
conditions where a low level of contamination is present.
 The following are liquid chemicals used for liquid sterilization includes:
I. Hydrogen peroxide

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  Hydrogen peroxide is a liquid chemical sterilizing agent which is a strong oxidant and can
destroy extensive range of microorganisms.
 It is useful in the sterilization of heat or temperature-sensitive equipment like endoscopes.
In medical applications, a higher concentration (35-90%) is used.
 H2O2 has a short sterilization cycle time as these cycles are as short as 28 minutes where
ethylene oxide has cycles that as long as 10-12 hours.
 Though, hydrogen peroxide has Limitation like low material compatibility, lower capacity
of penetration, and associated health risks.
 Vaporized hydrogen peroxide (VHP) is used to sterilize mainly enclosed and sealed areas,
like entire rooms and aircraft interiors.
II. Glutaraldehyde
 Glutaraldehyde is an accepted liquid sterilizing agent which requires moderately long
immersion time. For the removal of all spores, it takes as long as 22 hours of immersion
time.
 The presence of solid particles increases the immersion time.
 The penetration power is also not enough as it takes hours to penetrate a block of tissues.
 The use of Glutaraldehyde is thus partial to definite surfaces with less contamination.
III. Hypochlorite
 Hypochlorite solution which is also called liquid bleach is another liquid chemical that can
be used as a disinfectant, even though sterilization is difficult to obtain with this chemical.
 Submerging devices for a short period in liquid bleach may kill some pathogenic organisms
but to reach sterilization submersion for 20-24 hours is compulsory.
 It is an oxidizing agent and thus acts by oxidizing organic compounds which results in the
alteration of proteins in microbes which might eventually lead to death.
 Suitable concentrations of hypochlorite can be used for the disinfection of workstations and
even surfaces to clean blood spills and other liquids. (Sapkota, May 17, 2020)

Refrigerating system in hospital

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The complexity of the services offered in specialized hospitals and clinics is associated with the
use of increasingly more modern equipment and techniques, as well as the creation of new
medicines.

The refrigerator contains different components. Some of them are the following.

 Compressor
 Condenser
 Expansion valve
 Evaporator
 Thermostat
 Interconnecting tubing.
A thermostat controls the refrigerator temperature.
Principles of operation

Refrigerant leaves the evaporator as gas at a low temperature and pressure. The
compressor builds up a pressure difference in the system, sketch refrigerant gas through a
suction valve, and circulating it to the condenser. Compressed gas enters the condenser at a
higher-than-ambient temperature and is cooled to a liquid. As the liquid refrigerant leaves the
condenser, a capillary tube or an expansion valve controls its flow to the evaporator. The
capillary tube forms a heat exchanger to help further cool the refrigerant. As the
refrigerant leaves, it enters a low
pressure area that permits it to expand quickly and evaporate, absorbing high
temperature from the refrigerator storage area, thereby cooling
the storage area and its contents. Finally, the refrigerant gas is circulated from the
evaporator back to the compressor and is drawn through the compressor
suction valve to repeat the cycle.

In hospital there are different types of refrigerating system. Some of them are listed below.

 Pharmacy refrigerating system

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  Laboratory refrigerating system
 Blood refrigerating system

Pharmacy refrigerating system

Properly preserving drugs, vaccines, serum, blood, tissues, organs for transplant and many other
items is necessary to ensure the best conditions for treatments and preventive activities in the
medical field. Refrigeration and climate control systems and equipment play a fundamental role
in this task. This opens up great opportunities for those working in the area but requires
knowledge and great responsibility.

There are different types of pharmacy refrigerators one them is dual sensors pharmacy/vaccine
refrigerator.
The Twin Sensors drugstore refrigerator is appropriate intendedfor chemist's shop, pharmacy co
mpanies, healing centers, widespread prevention centers to store medicines, testing kit,
reagents, immunizations etc. Dual sensors, one air sensor for rapid control response, one manufa
ctured goods sensor with glycerin bottle for samples imitation temperature put on
view, stored inside the refrigerator.

Lab Refrigeration

The refrigerator in a laboratory is one of the most important equipment. Its function is to
maintain, in a controlled environment (refrigerated space), various fluids and substances, so that
they are kept in good condition the lower the temperature, the lower chemical and biological
activity.

Managing temperatures inside the hospital laboratory surroundings isa process that requires cons
ideration and carefulness in order to positively affect operation. Appropriate refrigeration of
reagents, microbiological materials, blood products, urine, etc is essential to keeping up the
integrity and stability of these elements.

Towards facilitate thisperpetual process for monitoring and maintaining temperatures in laborato
ry refrigeration devices is needed.

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Quality standards for laboratory refrigeration devices are executing an automated and self
documenting temperature observing organization in order to correctly preserve consistent temper
atures within particular ranges.

Refrigerator of blood in bank

Blood bank refrigerators are the optimal choice for storing your blood samples safely. Blood is
an essential human resource and its importance for research purposes and medical treatment as
well as its storage conditions cannot be emphasized enough. Blood storage needs a high level of
temperature uniformity and reliable technology, which our refrigerators ensure.

Our blood bank refrigerators include features such as uniform fan cooling that circulates the air
around the stored blood. Polyurethane (CFC-free) insulation in cabinet walls together with
double-pane glass doors reduce the adverse effects of the ambient environment temperature and
promote temperature uniformity in the cabinets.

Whether a blood sample is donated or taken as part of a treatment, it is a human resource that
must not be spoiled and the safety of the samples is of highest priority. An unexpected
breakdown of your blood bank refrigerator may cause severe damage to your whole project and
research.

The two separate and independent cooling systems ensure that in the unlikely event of a
malfunction to one of the cycles, the other will take over and serve as a backup. The temperature
will be maintained, and your samples will be intact. (https://www.arctiko.com/products/plus4-c-
blood-bank-refrigerators/)

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1. Refrigerant
It is also called as coolant and is the working fluid for the refrigerator. It takes the heat from inside of the refrigerator and transports it to the outside. Most commonly used refrigerant in the
refrigerator are isobutane (used in modern fridges), CFCs and ammonia (toxic gas and not used in modern fridge)

2. Compressor
It circulates the refrigerant during the working of the fridge. It compresses the refrigerant gas and increases its temperature and pressure. Compressor is the heart of the fridge, without it its
working is not possible.

3. Condenser Coil
It is present outside at the back of the refrigerator. It has grill tube and looks like radiator. Its main purpose is to cools the hot and high pressure gases from the compressor. When the hot gases
pass through the condenser coil, it gets cool down by the cool air of the room and gets converted into high pressure liquid.

4. Expansion Device or Throttling Device

As its names indicated it expands the high pressure liquid refrigerant and reduces its temperature and pressure. The temperature drops to 20 degree Celsius and pressure to 0.6 bar.

5. Evaporator Coil
Evaporator coil is present inside the fridge. It takes the heat from the air inside the fridge and makes it cool. And this cool air inside the fridge takes the heat from the food materials and lowers
their temperature.

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REFERENCE
Methods of sterilization . (Ninth Edition, 2019). The International Pharmacopoeia .

Butler, G. (August 2009). Steam Sterilization Cycles. STERIS .

https://www.arctiko.com/products/plus4-c-blood-bank-refrigerators/. (n.d.). Retrieved 04 26, 2021,

from ARCTIKO.

Methods of sterilization. (2019). The International Pharmacopoeia .

Pascale Choquenet, S. D. (2017). STERILIZATION IN HEALTH CARE FACILITIES. Sterilization Guideline .

Sapkota, A. (May 17, 2020). Chemical methods of sterilization.

Tankeshwar, A. (April 24, 2020). Radiation Sterilization: Types, Mechanism, and Applications.

William A. Rutala, D. J. (2016 Aug 9). Disinfection and Sterilization in Health Care Facilities. eRA website.

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