DISINFECTION & DECONTAMINATION (YT VIDEO FOR REFLECTION PURPOSES) – LESSON 2 – PMLS LEC
DISINFECTION AND DECONTAMINATION OF
WORKING AREA
o This lesson is an essential aspect of making
our surroundings clean and free from
contaminants.
o Contaminants are very common in any
working area inside the laboratory. Dirt is easy
to detect but biohazards are not.
o Therefore, a thorough discussion about
disinfection will introduce us to the different
methods and disinfectants that can be used to
eliminate contaminants not only inside the
laboratory but also at home.
DEFINITION OF TERMS:
o High-touch surfaces – high-touch surfaces are
those that have frequent contact with hands.
(Doorknobs, call bells, bed rails, light switches,
wall area around the toilet, and edges of
privacy curtains)
o Low-touch surfaces – surfaces that have
minimal contact with hands. (Walls, ceilings,
mirrors, windows sills)
o Bactericidal – agents that usually kill target
organisms
o Bacteriostatic – agents that inhibit bacterial
growth but generally do not kill the organism.
DISINFECTION AND DECONTAMINATION
o The process of removing or neutralizing
contaminants that have accumulated on the
workplace, personnel, and equipment.
o Intended to inactivate the microbes by
physical, chemical, and biological processes.
o inactivation process includes:
 proteins (structural proteins,
transport proteins, enzymes)
 nucleic acids (genomic DNA or RNA,
mRNA, tRNA, etc.)
 lipids (lipid bilayer membranes, other
lipids)
ANTISEPTICS VS DISINFECTANTS
ANTISEPTICS
 used on skin and mucous membranes to kill
microorganisms.
 Not for use on inanimate objects.
 Ex: hand sanitizers
DISINFECTANTS
 Use to kill microorganisms on inanimate
objects
 Not for use on skin or mucous membranes
 Ex: Mr. clean
PROPERTIES OF AN IDEAL DISINFECTANT
1. Wide spectrum of antimicrobial activity
2. Not affected by the environmental factors
3. Relatively safe to people that may be exposed
4. Should be stable
5. Odorless
6. Surface compatibility
7. Contact time and temperature
8. Residual activity and effects on fabric and
metal
9. Relatively inexpensive
10. Environment friendly
METHODS OF DISINFECTION
o Has 2 general methods: physical and chemical
method
PHYSICAL METHOD
The three physical methods of disinfection are:
1. Boiling at 100 degrees Celsius for 15 minutes,
which kills vegetative bacteria
2. Pasteurizing at 63 degrees Celsius for 30
minutes or 72 degrees Celsius for 15 seconds,
which kills food pathogens without damaging
the nutritional value or flavor.
3. Using non-ionizing radiation such as
ultraviolet (UV) light
o UV rays are long wavelengths and low energy.
They do not penetrate well, and organisms
must have direct surface exposure, such as
the working surface of a BSC, for this form of
disinfection wot work.
CHEMICAL METHOD
Chemical disinfectants comprise many classes
including:
DISINFECTION & DECONTAMINATION (YT VIDEO FOR REFLECTION PURPOSES) – LESSON 2 – PMLS LEC
Alcohols
o Refers to two water-soluble chemicals:
ethyl alcohol and isopropyl alcohol
o These alcohols are rapidly bactericidal
rather than bacteriostatic against
vegetative forms of bacteria
o They are not effective against bacterial
spores and have limited effectiveness
against non-enveloped virus.
o Higher concentration are less effective as
the action of the denaturing proteins is
inhibited without presence of water.
Aldehydes
o Formaldehyde is the main aldehyde used for
disinfection
o Formaldehyde in solution is useful for
decontaminating certain instruments
o Disadvantage: irritating fumes; not generally
used for surface disinfectants.
Halogens
o Halogens, especially chlorine and iodine are
frequently used as disinfectant
o Chlorine is most often used in the form of
sodium hypochlorite (NAOCl), the compound
known as household bleach.
Heavy Metals
o The most widely used heavy metals are those
mercury, silver, and copper
o High concentration of heavy metals
coagulates cytoplasmic proteins, resulting in
the damage or death of cell.
Quaternary ammonium compounds (QUATS)
o Used to disinfect benchtops or other surfaces
in the laboratory.
o Disadvantage: surfaces grossly contaminated
with organic materials, such as blood, may
inactivate quaternary ammonium compounds,
thus limiting their utility.
Phenolics
o Phenol disinfectants are effective against
bacteria (especially Gram-positive bacteria)
and enveloped viruses.
o Ex: Benzyl-4-chlorophenol, amyl phenol,
phenyl phenol
o 3% phenolic detergent can be used to clean
laboratory glassware
o Advantages: good general-purpose
disinfectant, not readily inactivated by organic
matter, active against wide range of
organisms
o Disadvantages: not sporicidal; can only be
achieved unless the items are immersed.
Hypochlorite
o Most widely used chlorine disinfectants and
are available in a liquid (e.g., sodium
hypochlorite) or solid (e.g., calcium
hypochlorite) form.
o The most common chlorine products are in
aqueous solutions of 4-6% sodium
hypochlorite, which are readily available as
household bleach.
o Advantages: broad spectrum of antimicrobial
activity, unaffected by water hardness,
inexpensive, fast acting, low incidence of
serious toxicity.
o Disadvantages: corrosive to metals at high
concentration (>500 ppm), inactivation by
organic matter, discoloring or bleaching of
fabrics, and release of toxic chlorine gas when
mixed with ammonia or acid.
Iodine
o Prepared either as a tincture with alcohol or
as an iodophor coupled to a neutral polymer
(e.g., povidone-iodine)
o 70% ethyl alcohol, followed by an iodophor is
the most common compound used for skin
disinfection before drawing blood specimens
for culture or surgery.
o Used as an antiseptic and disinfectant for
blood culture bottles and medical equipment
such as thermometers and endoscopes.
Hydrogen peroxide
o Used as antiseptics to clean wounds, cleaning
surgical sites after closure but can prolong
healing time if used in high concentration.
o Stabilized hydrogen peroxides can be used as
disinfectant for environmental surfaces.
Detergents and soaps
o Widely used as surface active agents, wetting
agents, and emulsifiers
o The most important antibacterial agents:
cetrimide and benzalkonium chloride
o Saturated fatty acids are more effective
against gram negative bacilli
DISINFECTION & DECONTAMINATION (YT VIDEO FOR REFLECTION PURPOSES) – LESSON 2 – PMLS LEC
o Unsaturated fatty acids have greater action
against gram positive.
MODE OF ACTION OF DISINFECTANTS
o Alteration of membrane permeability
o Damage to protein
o Rupture of cell membrane
o Damage to nucleic acids
o Interfere with metabolic pathway
FACTORS AFFECTING DISINFECTION
1. Types and number of microorganisms present
o Time necessary for killing microorganisms
increases in direct proportion to the number
of organisms (microbial load)
2. Temperature
o The lethal effect on the bacterial population
can be increased by increasing the
temperature.
3. pH
o A change of pH during the disinfection process
can affect the rate of growth inoculum
4. Concentration of disinfectant
5. Amount of organics present (blood, mucus, pus)
6. Nature of surface to be disinfected (e.g., potential
for corrosion, porous or nonporous surface)
7. Type of water available (hard or soft)
PRINCIPLES OF ENVIRONMENTAL CLEANING AND
DISINFECTION
o Cleaning helps to remove pathogens or
significantly reduce their load on
contaminated surfaces
o Cleaning with water, soap and some form of
mechanical action (brushing and scrubbing)
remove and reduces dirt, debris, and other
organic matter such as blood, secretions, and
excretions, but does not kill microorganisms.
o Organic matter can impede direct contact of a
disinfectant to a surface and inactivate the
germicidal properties or mode of action of
several disinfectants
o Disinfectant concentration and contact time
are also critical for effective surface
disinfection
o A chemical disinfectant, such as chlorine or
alcohol, should be applied after cleaning to kill
any remaining microorganisms
o Cleaning should progress from the least soiled
(cleanest) to the most soiled (dirtiest) areas,
and from the higher to lower levels so that
debris may fall on the floor and is cleaned last
in a systematic manner to avoid missing any
areas.
THE USE OF CHLORINE-BASED PRODUCTS
o Hypochlorite-based products include liquid
(sodium hypochlorite), solid or powdered
(calcium hypochlorite) formulations
o Hypochlorite displays a broad spectrum of
antimicrobial activity and is effective against
several pathogens at various concentrations
o 0.1% (1000 ppm) inactivates most other
pathogens that may be present in the health
care setting
o 0.5% (5000 ppm) is used for blood and body
fluids large spills
o Hypochlorite is rapidly inactivated in the
presence of organic materials
o Side effects: corrosion of metal, irritation of
skin or mucous membranes, chlorine smell
for vulnerable people like people with
asthma
o Chlorine solutions should be stored in
opaque containers, in a well ventilated
covered area that is not exposed to direct
sunlight
o Chlorine solutions are most stable at high pH
(>9) but the disinfectant properties of
chlorine are stronger at lower pH (<8)
o Solutions of 0.5% and 0.05% chlorine have
been shown to be stable for more than 30
days at temperatures of 25-35°C when the
pH is above 9.
DISINFECTION & DECONTAMINATION (YT VIDEO FOR REFLECTION PURPOSES) – LESSON 2 – PMLS LEC

shielded by objects, folded fabrics or surfaces

with intricate designs
o Ideally, chlorine solutions should be freshly
prepared every day.
HEALTH-CARE SETTINGS ENVIRONMENT

 Particular attention should be paid to

CALCULATION OF SODIUM HYPOCHLORITE
environmental cleaning of high touched
CONCENTRATIONS
surfaces and items: light switches, bed rails,
[% chlorine in liquid sodium hypochlorite / % chlorine door handles, intravenous pumps, tables,
desired] - 1 = Total parts of water for each part water/beverage pitchers, trays, and sinks
sodium hypochlorite.

NON-HEALTH CARE SETTINGS ENVIRONMENT

Ex: [5% in liquid sodium hypochlorite/ 0.5% chlorine
desired] -1 = 9 parts of water for each part sodium
hypochlorite
CALCULATION OF CHLORINE SOLUTIONS FROM
CALCIUM HYPOCHLORITE
 High-touch surfaces in these non-healthcare
settings should be identified for priority
disinfection. These include door and window
handles, kitchen and food preparation areas,
counter tops, bathroom surfaces, toilets and
taps, touchscreen personal devices, personal
computer keyboards, and work surfaces.

[% chlorine desired /% chlorine in hypochlorite

powder or granules] x 1 000 grams of calcium
hypochlorite powder for each litre of water.

Ex: [0.5% chlorine desired/35% in hypochlorite

powder] x 1 000 0.0143 × 1 000 = 14.3

Therefore, you must dissolve 14.3 grams of calcium

hypochlorite powder in each litre of water used to
make a 0.5% chlorine solution.

SPRAYING DISINFECTANTS AND OTHER NO-TOUCH

METHODS

o Routine application of disinfectants to

environmental surfaces by spraying or fogging
(also known as fumigation or misting) is not
recommended by WHO for COVID-19.

o It would not reduce an infected person's

ability to spread the virus through droplets or
contact

o Spraying may not be effective in removing

organic material and may miss surfaces