STERILIZATION

David, John Philip

De Guzman, Francis Carlo
Layug, RD Pauline
Lopez, Rose Anne
Santiago, Christine Nicole

Sterilization
-destroying all living organisms in a culture medium
or in a gas
Methods of Sterilization

Heat (moist or dry)

Chemical Agents
Radiation
Mechanical (ultrasonic vibrations)
Filtration

STERILIZATION METHODS
Heat/Thermal
most widely used means of sterilization
*Not all organisms have identical death

kinetics
*Death rate for moist cells is higher than
dry cells
*Moist heat is more effective than dry heat
*Dry heat is used only for glassware or
heatable solid materials

STERILIZATION METHODS
Chemical Agents
- oxidizing/alkylating abilities

-not advisable for the sterilization of medium

-often used for disinfection
Radiation

- absorbed UV light leads to DNA damage to cell

death
UV for surfaces, x-rays for liquids
Impractical sterilization tools (costly/safety)

Mechanical

Sonic/ultrasonic waves can disrupt and

kill cells
Extracting intracellular constituents
rather than sterilization technique

Filtration

- Removal of microorganisms from air or

other gases

THERMAL DEATH KINETICS

The destruction of microorganism by heat is considered as

loss of viability not destruction.

Thermal death of micro organisms at a particular

temperature can be described by first-order kinetics:
- eq. 1

Where,
n, is

the number of viable organisms present,

t, is the time of the sterilization treatment
kd, is the reaction rate constant of the reaction,
or the specific death rate per time.

Integration of eq. 1
- Eq. 2

n/n0 =e-kdt

Kd can be assumed to follow the Arrhenius

equation:

The graphically equations (1) and (2) are represented as,

The relationship observed in the above graph would be found only with
the sterilization of a pure culture in one physiological form, under ideal
sterilization conditions.

Predictions from the kinetic description:

An

infinite time is required to achieve sterile

conditions.
After a certain time there will be less than one
viable cell present.

DESIGN CRITERION
Del factor a measure of the size of the job
to be accomplished

Based on the sterilization criterion calculated, the

sterilization unit can be designed

BATCH
STERILIZATION

MODE

Sparging

Electrical
Heaters

Circulating
constant
pressure

CYCLE

TOTAL DEL-FACTOR

Direct steam
sparging

Constant
rate of heat
flow

Steam
circulation
through
heating coil

DIRECT STEAM SPARGING

where T = absolute temperature, K

To = initial absolute temperature of medium, K
H = enthalpy of steam relative to raw medium
temperature, J/kg
ms= steam mass flow rate, kg/s
c = specific heat medium, J/kg-K
M = initial mass of medium in batch sterilizer,
kg
t = time, s

CONSTANT RATE OF HEAT FLOW

(ELECTRICAL HEATING)

where

q = rate of heat transfer, J/s

STEAM CIRCULATION THROUGH

HEATING COIL

Where
TH = absolute temperature of heat
source, K
U = overall heat transfer coefficient,
J/s-m2-K
A = surface area across which heat
transfer occurs
during
sterilization

Where h = individual heat transfer

coefficient (W/m2K)
k = thermal conductivity (W/mK)

Length of controlled holding

period

BATCH COOLING USING NON-ISOTHERMAL HEAT

SINK AS PASSING COOLING WATER THROUGH
COOLING COIL

Where
Tco= initial absolute temperature
of heat sink, K
mc= coolant mass flow rate, kg/s

TO OBTAIN

Plot kd as a function of time

Integrate

the equation

KD VERSUS TIME

THOLD

A fermenter containing 40m3 (V) of medium (25 deg C ,To) is going to be

sterilized by the direct injection of saturated steam. The typical bacterial count
of the medium is abou 5x1012 m-3(Cn), which needs to be reduced to such an
extent that the chance for a contaminant surviving the sterilization is 1 in
1000(n). the steam(345 Kpa, abs,P steam) will be injected with a flowrate of
5000 kg/hr(ms) which will be stopped when a medium temperature reaches
122deg C(T). During the holding time, the heat loss through the vessel is
assumed to be negligible. After a proper holding time, the fermenter will be
cooled by passing 100m3/hr of 20deg C(Tco) water through the cooling coil in
the fermenter until the medium reaches 30 deg C. The coil has a heat transfer
area of 40m2 and for this operation the average overall heat transfer coefficient
for cooling is 2500 KG/hr-m2-K. The heat resistant bacterial spores in the
medium can be characterized by an Arrhenius coefficient Kdo of 5.7x1039/hr
and an activation energy Ed of 2.834x105 KJ/mol. The heat capacity and
density of the medium are 4.187KJ/kg-K and 1000kg/m3. Estimate the required
holding time

A continuous sterilizer with a steam injector and a flash cooler will be

employed to sterilize medium continuously with the flow rate of 2m 3/hr.
The time for heating and cooling is negligible with this type of sterilizer.
The typical bacterial count of the medium is about 5x10 12 m-3 which needs
to be reduced to such an extent that only one organism can survive during
two months of continous operation. The heat resistant bacterial spores in
the medium can be characterized by an Arrhenius coefficient(kdo) of
5.7x1039/hr and activation energy Ed of 2.834x105 KJ/kmol. The sterilizer
will be constructed with pipe with an inner diameter of 0.102m. Steam at
600kPa is available to bring the sterilizer to an operating temperature of
125 deg C. The physical properties of the medium at 125deg C are
c=4.187kJ/kg-K, density= 1000kg/m3 and viscosity of 4kg/m hr.What length
should the pipe be in the sterilizer if you assume ideal plug flow?

A filter bed of glass fibers(Dc=15 micrometer, Bed depth B=10cm and packing
density =0.03) is being used to sterilize air (20deg C, 1 atm) with an
undisturbed upstream velocity of vo of 10cm/s. The air stream contains 5000
bacteria/m3 (dp=1 micrometer, density p=1g/cm) Estimate the single fiber
collection efficiency by mertial impaction, and interception.

Properties of Air
Density=1.205kg/m3
Viscosity=1.82x10-5 Pa-s