Moist heat

Readily kills viruses, bacteria, and fungi

• Micro-organisms can be exposed to moist heat by using hot water, boiling

water, steam at atmospheric pressure (steaming) or steam under pressure

• Destroy micro-organisms by causing protein coagulation or denaturation

• Kill micro-organisms at lower temperatures and in shorter times than dry heat
 Saturated steam

• Steam produced at temperature

corresponding to the liquid boiling
point appropriate to its pressure

Phase diagram
 To summarize, why saturated steam is advantages

1. It flows quickly to and if required into every article in the load

(Volume contraction)

2. It rapidly heats the load to sterilisation temperature (liberation

of latent heat)

3. It provides, at high temperature, the moisture essential for

killing micro-organisms (production of condensate)
 Superheated steam

Superheated steam, steam is no longer

at the temperature corresponding to the
liquid boiling point appropriate to its
pressure and is called superheated
steam

Very slight cooling will not make super-

heated steam to condense
Phase diagram
 Steam for sterilisation

Portable sterilisers Large sterilisers (Dry saturated steam)

(Wet saturated steam) For surgical dressings and batches
intravenous fluids

Superheating is unlikely ✓ Superheating can take place in

several ways
 Superheating can take place in several ways

1. If the jacket temperature is higher than the chamber temperature

2. If Air is mixed with steam in the chamber

3. As the steam passes through the reducing valve

4. By the heat produced when cellulosic fibres absorb moisture

 Superheating can take place in several ways

1. If the jacket temperature is higher than the chamber temperature

The body of the large steriliser is surrounded by a steam jacket.

To prevent excessive condensation on the inner
surface of the chamber

Steam jacket is cooler Steam jacket is hotter

Condensate would soak dressings and Superheated steam

considerably prolong the drying stage
at the end of the process

Therefore, the jacket and chamber should be at the same temperature

 Superheating can take place in several ways

2. If Air is mixed with steam in the chamber

• If at 2 bars, the chamber contains 10% of air this will contribute 0.2
bars of the total pressure and only 1.8 bars will be due to steam

• Steam of this pressure will be saturated only at 117C and will be

superheated at the temperature corresponding to 2 bars at 121C
 Superheating can take place in several ways

2. If Air is mixed with steam in the chamber

• This superheated steam will not liberate its latent heat until it has been
cooled 4C

• This type of superheating is very difficult to detect, so air must be removed as

completely as possible from steriliser heated by steam alone
 Superheating can take place in several ways

3. As the steam passes through the reducing valve

• Pressure of the steam supply (6.3 bars) is usually much greater than the
pressure required for sterilization

• It must be passed through a reducing valve before admission to

steriliser. The heat content is unaffected, steam becomes super heated

• But this rarely happens in practice

• As steam arriving at the valve contains suspended water droplets and

excess heat is completely absorbed by evaporation of these
 Superheating can take place in several ways

4. By the heat produced when cellulosic fibres absorb moisture

• If materials have very low moisture content, absorb of moisture and

evolution of heat continue after the steam temperature has been reached
and consequently, the fibres and near by steam becomes superheated

• Correctly stored fabrics and dressings contain 5% of moisture, and the

few degrees of superheating can be tolerated

• Storage under very dry conditions can lead to excessive superheating

and, therefore, prevent sterilisation
 Superheating can take place in several ways

4. By the heat produced when cellulosic fibres absorb moisture

• If the sterilizer containing dressings is preheated by the jacket for some

time before steam is admitted the fibers become very hot as well as very
dry

• On admission of steam, heat absorption is added to the heat already in

fibres and considerable superheating takes place it may be at 50C or
more. It sometimes causes dressings to burst into flames when the
chamber door is opened
 Tolerable degrees of Superheating

• If steam is superheated it would be not expected to be satisfactory

sterilising agent

• However, Savage (1937), 5 to 15 or even more degrees of superheat,

depending on the saturation temperature before superheating, can be
tolerated

• Most sensible procedure is to prevent superheating in every possible

way and to set the tolerable maximum low e.g 5C
Applications

1. Glass Apparatus and containers: It is better to use dry heat for the sterilization
of the containers and apparatus made entirely of glass because with moist
heat----
i. Post –sterilization drying is necessary
ii. Air drainage must be ensured by suitable loading
iii. Extraction from and damage to the glass surfaces are more likely

2. Normally, dry heat is used for loosely fitting metal caps but metal or
thermostable plastic screw caps with rubber liners must be autoclaved

Rubber closures for multi-dose containers are most conveniently autoclaved in

the solution with which they have been equilibrated
Applications

3. Injection solutions and suspensions: There are more than 100 official
injections and the majority are sterilized by autoclaving

4. Surgical dressing and fabrics: cotton-wool balls, gauze swabs, ribbon

gauze, bandages, operating gowns, caps and masks, towels, trolley cloths, and
rubber sheeting

Successful sterilization requires careful selection of containers or packaging

material, faultless methods of packing, and correct loading in the sterilizer
 Advantages of autoclaving

1. Autoclaving destroys micro-organisms more efficiently than dry heat and,

therefore a shorter exposure at a lower temperature is possible

2. It can be used for a large proportion of the official injections

3. In a sterilizer supplied with dry saturated steam, porous materials can be

sterilized without damage

4. Equipment or components of rubber and certain plastics, such as nylon

and P.V.C., will withstand the conditions
Disadvantages of autoclaving

1. It is unsuitable for anhydrous materials (Powders and oils)

2. Injections and articles that deteriorate at 115 C

Testing the efficiency of sterilisers
 Steam steriliser

Regular tests-efficiency, working satisfactorily, and being operated correctly

Two types tests

1. Direct tests 2. Indirect tests

(sterility tests on the products) Instrumental
Cultural
Chemical
 1. Direct tests (sterility tests on the products)

Here, if the sterility of each load is proved by simple, rapid, and non-destructive
tests on every item before it was used-not possible

Many issues
• Highly skilled personnel
• Restricted choice of media and incubation conditions
• Accidental contaminations
• Not possible to test every item

This is not suitable method of testing a sterilizer

 2. Indirect tests (Instrumental)

If we can show satisfactory physical conditions for adequate time –then assume that
contents will be sterile

Ideal conditions include

• Temperature and time (More important)
• Vacuum (correct functioning of vacuum switch)
• Superheating was not excess (efficient air discharge)
 2. Indirect tests (Instrumental)

Temperature can be recorded easily

Thermometer (not sensed from inside the load)
Thermocouples inside dummy packs (More expensive)-The outputs from these can be
used to operate several recorders-This equipments is more expensive but gives very
quick responses to temperature changes

Faults will be detected- there will be no danger of issuing unsterile products

 2. Indirect tests (cultural)

Preparations containing bacterial spores dummy packs in the load tested for sterility
after exposure

Issues
Selection of spores
Sterility testing

Selection of spores

Resistance
Bacillus subtilis (Unwise choice-killed in few minutes in boiling water) and
dangerous for routine use
 2. Indirect tests (cultural)

• Soil samples or thermophilic bacteria (Choices)

• Soil samples

• Complicated due to soil particles

• Subculturing will be necessary (unsuccessful, if the organisms need growth

factors from the soil particles)

• Soil organisms should be wetter than the organisms in or on the article

 2. Indirect tests (cultural)

Thermophilic bacteria

Advantages
Cultivation and spore production can be controlled
Non-specific turbidity-low
Definite number of spores- test paper

Spores of Bacillus stearothermophillus are used-moist heat (5 min at 121℃)

Spores of Clostridium tetani, Bacillus stearothermophillus – Dry heat

recommended
 2. Indirect tests (cultural)
Sterility testing
Thick filter paper strips- impregnated with the spores and usually they are sterilized
inside glassine envelopes (glossy transparent paper)

Precautions

The medium must satisfy the nutritional requirements

Incubation conditions

Several strips should be placed in each site in the sterilizer to prevent an incorrect
conclusion based on a paper of unusual resistance

Removal of strips should be careful

 2. Indirect tests (Chemical)

• Types that cannot indicate a satisfactory exposure

• Types that indicate a satisfactory exposure

 2. Indirect tests (Chemical)

Types that cannot indicate a satisfactory exposure

• Witness tubes
• Klintex papers
• Test tablets
• Bowie-dick test
Witness tubes

Sealed glass tubes containing a chemical that melts at sterilization temperature

(acetanilide at 115℃ and benzoic acid at 121℃)- gave no indication of the time
exposure

A dye such as methylene blue which gave a different colour when dissolved in the
melt, was included
 2. Indirect tests (Chemical)
Advantages over witness tubes and not affected by dry heat
Klintex papers
Test tablets

Klintex papers (Paper strips with coloured coating that disappears in presence
of steam-leaves AUTOCLAVED)-even 2 min boiling water can do this
The strips are coated with a thermochromic ink that reacts to moist heat.
Test tablets
75% lactose, 24% starch and 1% magnesium trisilicate

They are hard and white at first but autoclaving-Brown gelatinous (24 min at 115℃)-
time of exposure is not monitored
 2. Indirect tests (Chemical)

Bowie-dick test

• High vacuum autoclave-to make sure all that steam has displaced all the air
from the porous load

• A test pack is made from huckaback towels 1 X 0.6 m in size, folded into 8
thicknesses-give you a pile of 0.25 m high

• Center of the pile- sheet of paper with high permeability to steam to which two
pieces of autoclave tape have been stuck from corner to corner in the shape of a
cross
 2. Indirect tests (Chemical)

Bowie-dick test

• Greater exposures may change the tape even when air removal is inadequate

• Packed and placed in a cardboard box- steriliser-based on color changes

throughout the strips if steam has fully penetrated the pack

• If air remains, the bars in the centre are lighter in colour

References

• Hugo& Russell’s Pharmaceutical Microbiology 8th Edition

• Cooper and Gunn’s Dispensing for pharmaceutical students
12th Edition