STERILIZATION AND

DISINFECTION

09/19/16

STERILIZATION AND
DISINFECTION
STERILIZATION
The killing or removal of all
microorganisms including bacterial
spores
Autoclaving: exposure to steam at
121 C, 15 lb/in2 for 15 minutes
Ethylene oxide gas: if damaged by
moist heat
Filtration: intravenous solutions

STERILIZATION AND
DISINFECTION
DISINFECTION
The killing of many, but not all,
microorganisms
Pathogens must be killed, but
some organisms and bacterial
spores may survive.
Agents vary in their tissuedamaging properties.

STERILIZATION AND
DISINFECTION
DISINFECTION
Antiseptics: chemicals used to
kill microorganisms on the
surface of the skin and mucous
membranes

Rate of Killing of
Microorganisms
Two variables:
1. Concentration of the killing agent
2. Length of time it is applied
The number of survivors (N) is
inversely proportional to the
concentration of the agent (C), and
the time of application of the agent
(T).

CHEMICAL AGENTS
Phenol coefficient: a quantitative
measure of the ability of a
chemical to kill microorganisms.
- The ratio of the concentration
of phenol to the concentration of
the agent required to cause the
same amount of killing under
standard conditions of the test

CHEMICAL AGENTS
3 mechanisms:
1. DISRUPTION OF CELL
MEMBRANES
2. MODIFICATION OF PROTEINS
3. MODIFICATION OF DNA
Some chemicals act by more then
one mechanism.

A. Disruption of Cell
Membranes
1. ALCOHOLS
Ethanol: used to clean the skin
prior to immunization or
venipuncture
Disorganizes the lipid structure in
membranes
Denatures proteins
Requires the presence of water for
maximal activity.

A. Disruption of Cell
Membranes
2. DETERGENTS
Hydrophobic portion: lipid-soluble
Hydrophilic portion
Surfactants interact with the lipid
in the cell membrane and with the
surrounding water
Quaternary ammonium compounds
e.g. benzalkonium chloride (skin
antiseptic)

A. Disruption of Cell
Membranes
3. PHENOLS
Original disinfectant used by
Lister (1860s)
Rarely used today, too caustic
Hexachlorophene is used in
germicidal soaps (Phisohex)
- Possible neurotoxicity
Cresol (methylphenol): the active
ingredient in Lysol
Also denatures proteins

B. Modification of
Proteins
1. CHLORINE
Used to purify the water supply
and to treat swimming pools
Active component of
hypochlorite (bleach, Clorox)
An oxidizing agent; cross-links
sulfhydryl groups in enzymes to
form an inactive disulfide)

B. Modification of
Proteins
2. IODINE
The most effective skin
antiseptic used in medical
practice. Two forms:
Tincture of iodine (2% solution
of iodine and potassium iodide
in ethanol
- Irritating to the skin

B. Modification of
Proteins
2. IODINE
Iodophors: complexes of iodine
with detergents
- Used to prepare the skin prior to
surgery
- Prior to blood culture
- Less irritating
Acts like chlorine
Also binds specifically to tyrosine
residues in proteins

B. Modification of
Proteins
3. HEAVY METALS
Mercury and Silver
Bind to sulfhydryl groups, blocking
enzymatic activity
Merthiolate and Mercurochrome
Silver nitrate drops: prevent
gonococcal ophthalmia
neonatorum
Silver sulfadiazine: prevent
infection of burn wounds

B. Modification of
Proteins
4. HYDROGEN PEROXIDE
Used to clean wounds and to
disinfect contact lenses
Limited by the organisms ability to
produce catalase
Bubble formation: oxygen arising
from the breakdown of H2O2 by tissue
catalase
An oxidizing agent; attacks
sulfhydryl groups, inhibits enzyme
activity

B. Modification of
Proteins
5. FORMALDEHYDE and
GLUTARALDEHYDE
Formaldehyde (Formalin = 37%
solution in water)
- Denatures proteins and nucleic
acids
Glutaraldehyde
- Ten times more effective than
formaldehyde and is less toxic
- Used to sterilize respiratory therapy
equipment

B. Modification of
Proteins
6. ETHYLENE OXIDE GAS
For sterilization of heatsensitive materials
Kills by alkylating both proteins
and nucleic acids;
The hydroxylethyl group attacks
the reactive atoms on essential
amino and hydroxyl groups

B. Modification of
Proteins
7. ACIDS and ALKALIS
Strong acids and alkalis
denature proteins
Weak acids are used as food
preservatives; are bacteriostatic
- e.g. benzoic, propionic, citric
acid

B. Modification of Nucleic
Acids
CRYSTAL VIOLET (gentian violet)
Skin antiseptic
Binding of the positively charged
dye molecule to the negatively
charged phosphate groups of the
nucleic acids
MALACHITE GREEN
Lowenstein-Jensens medium

PHYSICAL AGENTS
HEAT
Denatures proteins
Membrane damage and
enzymatic cleavage of DNA may
also be involved
Moist heat: lower temperature;
water aids in the disruption of
covalent bonds

PHYSICAL AGENTS
HEAT
Moist Heat Sterilization (autoclaving)
Most frequently used in Medicine
Most bacterial spores are resistant
to boiling
Steam; 15 lbs/in2 , 121 C; 15 20
minutes
Dry Heat Sterilization
180 C for 2 hours
Glassware

PHYSICAL AGENTS
HEAT
Pasteurization
Heating milk to 62 C for 30
mins., followed by rapid cooling
Flash Pasteurization: 72 C for
15 seconds
Kills milk-borne pathogens

PHYSICAL AGENTS
RADIATION 2 types:
1. Ultraviolet light
Wavelength region of maximum
absorption by purine and pyrimidine
bases of DNA
Formation of thymine dimers
Also addition of hydroxyl groups to
the bases
DNA replication is inhibited
Can damage the cornea and the skin

PHYSICAL AGENTS
RADIATION 2 types:
2. X-rays
Higher energy and penetrating power
vs. ultraviolet light
Production of free radicals e.g.
production of hydroxyl radicals by
the hydrolysis of water; breaks
covalent bonds in DNA
Also direct hit on covalent bonds in
DNA

PHYSICAL AGENTS
RADIATION 2 types:
2. X-rays
Spores are resistant to X-rays
due to low water content
For sterilization of heatsensitive items

PHYSICAL AGENTS
FILTRATION
For sterilizing solutions with heatsensitive components
Intravenous solutions are filtered
before autoclaving
Physically traps particles larger
than the pore size
Retains smaller particles by
electrostatic attraction of the
particles to the filter

PHYSICAL AGENTS
FILTRATION
Nitrocellulose: pore size retains
bacteria and spores

STERILIZATION AND
DISINFECTION

09/19/16