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Testing of Disinfectants

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40 views33 pages

Testing of Disinfectants

Uploaded by

dodeti8872
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Evaluation of Potency of

Disinfectants
To know required effective
dilution

Why to test a To know time taken for onset


of action
Disinfectant???
Periodical monitoring of disinfection
ability/activity as disinfectants can
loose their anti-microbial
activity/action on long standing and in
presence of organic matter
Principle

Expose the indicator bacteria to the disinfectant

Test the viability of indicator bacteria by sub-culturing them on media


prepared without disinfectant.

The disinfectant carried over to subculture can be neutralized by diluting them to sub-
inhibitor conditions or by adding inactivator like 0.5% Sodium thiosulphate for chorine
compounds, iodophores, 1% Sodium bisulphate for HCHO and glutaraldehyde
Bactericidal tests:

The test organism is exposed to a suitable concentration of the


disinfectant

Samples are taken at specified times and added immediately to a diluent


or culture medium containing the appropriate disinfectant inactivator

The treated samples are cultured for surviving microorganisms.


(Carrier test)
Phenol coefficient test

• This method is sometimes called FDA method or AOAC phenol-coefficient


(Association of Official Agricultural Chemists and Food and Drug
Administration).
• This is a good way to test disinfectants that are dissolvable in water.
• measure of the bactericidal activity of a chemical compound in relation
to phenol.
• This procedure uses a particular strain of Salmonella
typhi and Staphylococcus aureus as the test organism.
• There are two distinct methods of Phenol coefficient test, such as;
1. Rideal Walker method
2. Chick Martin test
Rideal Walker method for Phenol coefficient test

• Rideal and Walker developed this method of phenol coefficient


testing to test the effectiveness of disinfectants against phenol to kill
Salmonella Typhi.
Rideal Walker Test

• Phenol coefficient obtained by dividing the dilution factor at


which the tested substance shows activity by the dilution
factor at which phenol shows comparable activity.
• RW coefficient= Dilution of disinfectant killing in 7.5 min but
not in 5 min/Dilution of phenol killing in 7.5 min but not in 5
min
Advantages of the Rideal-Walker test

• This test is highly reproducible and provides a detailed description of


the coefficient.
• It is easy to do and it can be done quickly.
• It is possible to quantify the effectiveness of the disinfectant.
Disadvantages of Rideal Walker method
• Only Phenolic disinfectants can be tested
• No organic matter used
• Short Disinfection time
• Only one MO tested
10
Capacity Tests
• The ability to retain activity in the presence of increasing load is capacity of
disinfectant
• Disinfectant is repeatedly challenged by successive additions of bacterial suspension
until its capacity to kill has been exhausted
• Capacity test simulate the practical situations of housekeeping and instrument
disinfection.
• Kelsey-Sykes test: triple challenge test
• Designed to determine the conc of disinfectants in clean and dirty conditions
• Disinfectant challenged by 3 successive additions of bacterial suspension
• Takes over 30 min to perform
• Autoclaved yeast cells are added which reduce the conc of disinfection to half (final
conc 0.5%)
• Single test organism selected from Staphylococcus aureus, E. coli, Pseudomonas
aeruginosa, P. vulgaris
• Clean condition: Hard water; Dirty conditions: yeast suspension
Kelsey Sykeys Test

Reported as pass/fail test based on microbial growth


Stability Tests

• If Disinfectant is diluted and stored, its effectiveness should be confirmed by supplementary


stability tests
• Pseudomonas aeruginosa used as test microorganism
• Disinfectant is diluted, one portion inoculated immediately and other kept and inoculated
with test MO after 7 days
• Tested for growth for 7 days, If growth detected, higher conc of disinfectant should be used
Practical Tests
• After measuring the time-concentration
relationship for the disinfectant in a quantitative
suspension, practical tests are conducted under
real-life conditions.
• It is important to determine if the proposed use
of dilution is still appropriate under the
conditions in which it will be used.
• Surface Disinfection tests
Surface Disinfection tests

Assess the effectiveness of disinfectant against surface adhered MO.

The test surface is contaminated with Standardized inoculum of test bacteria and dried.

Then a definite volume of disinfectant solution is distributed over the


carrier.

After the given exposure time, the number of survivors is determined by impression on contact
plate or by a rinsing technique in which the carrier is rinsed in a diluent and the number of surviving
bacteria is determined in the rinsing fluid.
In use hard surface disinfection tests-
floors, walls, table tops
In-Use test
• Described by
Maurer in 1985
• Used in hospitals
and labs to detect
contamination of
disinfectants
Disk diffusion Method

• This involves using various chemicals to prepare sterile filter paper disks.
• Next, place the disks on an inoculated agar plate (seeded agar plate).
• The chemicals are released from the disks onto the agar.
• Clear areas around the disks are zones of inhibition as the bacteria “lawn”
grows.
• There are many factors that influence the size of the inhibition zones, such as
whether the agent can dissolve in water or diffuse well in the agar.
• However, larger inhibition zones usually correlate with greater inhibition
effectiveness.
• Each zone’s diameter is measured in millimeters.
Disk diffusion Method
Minimum Inhibitory concentration (MIC)

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