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Definition

STERILIZATION

CPB 30103
Biochemical Engineering

Law Jeng Yih

Sterilization Methods Sterilization by Heat

Sterilization by Heat Sterilization by Heat

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Sterilization by Heat Sterilization by Heat

Sterilization by Heat Sterilization by Heat

Sterilization by Heat Sterilization by Heat

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Sterilization by Heat Sterilization by Heat

 Autoclave

Sterilization by Heat Sterilization by Heat

Ultraviolet Light Ultraviolet Light

Sterilization Sterilization

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Sterilization by Filtration Chemical Sterilization

Chemical Sterilization Chemical Sterilization

Chemical Sterilization Thermal Death Kinetic

 The thermal death kinetics may be represented by the following equation:
-dN/dt = kd N (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.
 On integration of equation (i) from t=0’ to t=t,we have the following expression :
Nt/N0 =e-kdt (2)
 where
 No is the number of viable organisms present at the start of the sterilization
treatment,
 Nt is the number of viable organisms present after a treatment period, t.

On taking natural logarithms, equation (2) is reduced to:

ln(Nt/N0) = - kd t (3)

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Thermal Death Kinetic Thermal Death Kinetic

The graphically equations (2) and (3) are represented as, Initial population increase resulting from the heat activation of
spores in the early stages of a sterilization process
Fig: Plots of the proportion of
survivors and the natural
logarithm of the proportion of
survivors in a population of
microorganisms subjected to a
lethal temperature over a time
period.

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.
From Principles of Fermentation From Principles of Fermentation
Technology,- Peter F. Stanbury, Allen Technology,- Peter F. Stanbury, Allen
Whitaker, Stephen J. Hall, Second Whitaker, Stephen J. Hall, Second
Edition, Edition,

Thermal Death Kinetic Thermal Death Kinetic

An initial stationary period observed during a sterilization treatment Initial population decline at a sub-maximum rate during a sterilization
due to the death of spores being completly compensated by the heat treatment due to the death of spores being compensated by the heat
activation of spores activation of spores

From Principles of Fermentation From Principles of Fermentation

Technology,- Peter F. Stanbury, Allen Technology,- Peter F. Stanbury, Allen
Whitaker, Stephen J. Hall, Second Whitaker, Stephen J. Hall, Second
Edition, Edition,

Thermal Death Kinetic Thermal Death Kinetic

The effect of a sterilization treatment on a mixed culture consisting of a
The effect of a sterilization treatment on a mixed culture consisting
high proportion of a relatively resistant organism
of a high proportion of a very sensitive organism

From Principles of Fermentation From Principles of Fermentation

Technology,- Peter F. Stanbury, Allen Technology,- Peter F. Stanbury, Allen
Whitaker, Stephen J. Hall, Second Whitaker, Stephen J. Hall, Second
Edition, Edition,

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Thermal Death Kinetic Thermal Death Kinetic

The probability of an unsuccessful sterilization:
1- P0(t)
The probability of extinction (successful sterilization):
P0(t)=[1-p(t)]N0
where p(t) is the probability that an individual will still be viable
at time t. N0 is the number of individuals (cells or spores)
initially present.

Assume first order death rate:

p(t)=e-kdt
1- P0(t)=1-[1-p(t)]N0 = 1-[1-e-kdt]N0
where the specific death rate kd = αe-E0d/RT(1/time), constant at specified
T. Eod is the activation energy for the death of the organism.

Thermal Death Kinetic Thermal Death Kinetic

Decimal reduction time (D): time for the number of 1- P0(t)= 1-[1-e-kdt]N0
viable cells to decrease tenfold (N(t)/ N0 = 0.1). kd = αe-E0d/RT
From the above equation:
N(t) = N0 p(t) - Known N0, T, t, determine Kd, the probability of an
unsuccessful sterilization is determined.
N(t) = N0 e-kdt
0.1= e-kdD - Given N0, T, acceptable probability of failure e.g. 10-3,
required time can be determined
D=2.303/kd
- Higher Kd tends to achieve low probability of sterilization
N(t) is the expected value of the number of failure. Normally at 121oC.
individuals present at time t.
Kd of vegetative cells > 1010 min-1, spores 0.5-5 min-1. The
major concern is spores.

Difficulty of Sterilization
Sterilization Chart
Increases with Scale
e.g. N0=108,
Kd=1min-1 (1210C), t=26min
Kdt=26
Probability of failure:
1-P=0.001

e.g. N0=108,
Kd=1min-1 (1210C), 1-P=0.001
Kdt=26
Required sterilization time:
t=kdt/kd=26 min
(M. Shular, Textbook, p.319)

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Difficulty of Sterilization
Batch Sterilization
Increases with Scale
(Sterilization)
holding

T (K) Kd2

T (K)
Kd1 Kd3

t time

t time

Temperature verse time in a batch sterilization process

Batch Sterilization and Continuous versus Batch

Continuous Sterilization Sterilization

Comparison of a batch (A)

with a continuous (B)
sterilization for the
temperature profile of the
medium sterilized.
The continuous unit allows
short-time, high
temperature sterilization.