PRINCIPLES OF

STERILIZATION METHODS:
KILLING BY MEANS OF HEAT

Raditya Weka Nugraheni

 OUTLINE
• History
• Heat Inactivation Kinetics
• Moist Heat and Dry Heat
• Mechanism of Microbial inactivation
• Mechanism of Spore Resistance to heat
• Conclusion
HISTORICAL ASPECTS OF STERILIZATION
 The fumes of burning chemicals were
also used by the ancients for deodorizing
and disinfecting purposes. Of early
importance was sulfur, apparently the first
of the useful chemicals to be mentioned.
In the Odyssey, the following passage
may be found:

“To the nurse Eurycleia then said he:

Bring cleansing sulfur, aged dame, to me
And fire, that I may purify the hall.”

3,000 BC – The use of antiseptics such as pitch or tar, resins and

aromatics was widely employed by the Egyptians in embalming
bodies even before they had written language. They were also
acquainted with the antiseptic value of dryness resulting from
the use of certain chemicals such as niter and common salt.
They became so skillful in the art of embalming that mummies
of thousands of years old are still in good condition.
 Galen (130-200 AD), a Greek who
practiced medicine in Rome and was the
most distinguished physician after
Hippocrates, boiled instruments used in
caring for wounded Roman gladiators.
His writings and those of Hippocrates
were the established authority for
medicine for many centuries.

In Persia, Avicenna (980-1046) indicated in

Hippocrates of Cos (460-377 BC), Book III of his Canon that water may be
was the first to separate medicine rendered drinkable by evaporation and
from philosophy and disproved the distillation, or simply by boiling. Sayyid
idea that disease was punishment Ismail Jorjani (1042-1135) observed that
for sin. He also advocated filtered (or boiled) water took longer to go
stale
irrigation of wounds with wine or
boiled water, foreshadowing
asepsis.
 Charles Chamberland, Louis
Pasteur’s pupil and
collaborator, developed the
first pressure steam
sterilizer, or autoclave in
1876.

Louis Pasteur

Charles Chamberland
• The research of Robert Koch and his
associates in 1881 on the disinfecting
properties of steam and hot air mark the
beginning of the science of disinfection and
sterilization. They devised the first
nonpressure flowing steam sterilizer.
• 1881 – Sterilization by boiling was
introduced. Everything used during an
operation, including linens, dressings, and
gowns was boiled.
HEAT INACTIVATION KINETICS
 The goal of thermal processing for sterilization is to deliver
sufficient heat to inactivate all of the organisms that might be
present. While this is the ultimate objective, as a result of the
inactivation kinetics of microorganisms by heat (see below), a
more accurate definition is to reduce the probability of survival
of microorganisms to an acceptably low level.
 Initial Bacteria Remaining
Time Logarithm of
Bacterial Killed in 1 Bacterial
(mins.) Survivors
Count Minute Count

1 1,000,000 900,000 100,000 5

2 100,000 90,000 10,000 4

3 10,000 9,000 1,000 3

4 1,000 900 100 2

5 100 90 10 1

6 10 9 1 0

7 1 0.9 0.1 -1

8 0.1 0.09 0.01 -2

9 0.01 0.009 0.001 -3

Dr. Sugiyartono, 2015 10 0.001 0.0009 0.0001 -4

11 0.0001 0.00009 0.00001 -5

LOGARITHMIC MICROBIAL DEATH 12 0.00001 0.000009 0.000001 -6

Adapted from: Favero MS, Bond WW. chapter in Block SS, 5th Ed, 2001
1. The D-value (decimal reduction time (DRT)), is defined as the time in minutes at a particular constant
temperature to reduce the viable population by 1-log10, that is to 10% of the initial value, or by 90%.
2. The z-value is defined as the temperature (°C) to bring about a 10-fold reduction in D-value; it is obtained from the
slope of the curve in which the D-value on a logarithmic scale is plotted
against temperature on an arithmetic scale.
The F-value expresses a heat treatment at any temperature as equivalent to that effect
produced by a certain number of minutes at 121°C; F0 is the F-value when z is 10°C.
MOIST HEAT AND DRY HEAT STERILIZATION
 MOIST HEAT
• Sterilization in an autoclave using moist heat is optimal in saturated steam at the phase
boundary between the steam and condensate at the same temperature.
HOW AUTOCLAVES WORK?
 DRY HEAT
• Sterilization by dry heat is less efficient than by moist heat. Defnitions of D-value and z-value
given for moist heat above apply
equally here.
• Dry heat as a means of sterilization is reserved for those products and materials that contain little
or no water and cannot be
saturated with steam during the heating cycle (Table 15.1.5)
• Sterilization depends upon heat transfer from a
gas (hot air) to cooler objects and it is essential that even temperature distribution throughout the
sterilization chamber is
achieved. In practice, this is done by the inclusion of a fan unit
at the rear of the oven, which ensures forced air circulation.
 INACTIVATION OF BACTERIA
• There are several potential target sites in non-sporulating bacteria:
1. outer membrane of Gram-negative bacteria
2. cytoplasmic membrane,
3. ribonucleic acid (RNA) breakdown,
4. and protein coagulation
 CONCLUSION
• The various developments that aim to minimize heat damage to
the components of foods and pharmaceuticals, while at the same
time ensuring that the correct F0 is delivered, will probably remain
the most important targets in the near future.