Disinfectant

*Destruction (or) reduce (or) removal of micro organisms on non living organism (or)
inanimate (for example: Dishes, washing areas, benches, working areas).

*Terminology of Disinfectants:
o Sepsis: Bacterial contamination.
o Asepsis: Absence of contamination.
o Antisepsis: Chemical destruction of vegetative pathogen on living tissue.
o Sanitization: Lowering microbial count on eating or drinking utensil to safe level.
o Bactericidal: Chemical agents capable of killing of bacteria.
o Fungicidal: Chemical agents capable of killing of fungi.
o Viricidal: Chemical agents capable of killing of virus.
o Sporicidal: Chemical agents capable of killing spores.
o Bacteriostatic: The chemical agents inhibit the growth of bacteria.

 Disinfection generally kill the sensitive vegetative cells, but mot heat resistant
endospores.
 If object is inanimate (life less) or animate (living) human body / tissues disinfectants are
generally used in Bactericidal but occasionally bacteriostatic.

*Classification:
Chemical agents’ classification is acids and alkaline, alcohols, halogens, phenol and its
derivatives heavy metals, aldehydes quaternary ammonium compounds, dyes, detergents.

 Mode of action:
1. Alternation of membrane permeability.
2. Damage of protein.
3. Rupture of cell.
4. Damage of nucleic acid.
5. Interface with metallic pathway / metabolic pathway.

 Properties of disinfectant:
 Broad spectrum.
 Nontoxic.
 Fast acting.
 Odor less.
 Surface compatible.
 Economical.
 Easy to use.
 Soluble & miscibility.
 Stable on storage.
*Factors influencing the disinfectants:
o Concentration of disinfectants.
o Temperature.
o Time of contact.
o PH of environment.
o Surface tension.
o Formulation of disinfectant.
o Chemical structure of disinfectant.
o Types / number of microorganisms.
o Interfering with the substance.
o Potentiation, synergism, antagonism. Of disinfectants.
1) Concentration of disinfectants:
-Lethal effects of bacterial population increase by increasing in concentration of disinfectants.
-Effectiveness is generally related to concentration exponentially.
-Optimum concentration of phenol is 1%.
-This concentration is disinfecting effectiveness become less.
-The diluting co-efficient calculate from n = log t2 – log t1
log c2 – log c1
-n = Concentration exponent / dilution coefficient.
-log t2 = death time with disinfectant concentration c2.
-logt1 = death time with disinfectant concentration c2.

2) Temperature:
-Lethal effects of bacterial population increases by increasing in temperature.
-Effects of temperature on bactericidal activity may be expressed as quantitatively by means
of temperature coefficient.
-The temperature coefficient per degree raise in temperature is denoted as “Ø / Q10”.
- Ø = Time required to kill at Toc.
Time required to kill at T+100oc.

3)Time of contact:
-The sufficient time contact must be allowed for disinfectant to exhaust its action.

4)PH of environment:
-The PH is main factor for disinfectants.
-A change of PH during the disinfectant process can affect the rate of growth inoculum.
-PH 6.8 is optimal growth of bacteria and rate of growth declines on either side of range.
-The phenolic and acidic antimicrobial agents usually have greatest activity in acidic
conditions.
-ACIDINE DYES & quaternary ammonium compounds are usually a more active in alkaline
conditions.

5)Chemical structure of disinfectant:

-The chemical structure of compounds affects the disinfectant activity.
-Generally, Halogenations increases the anti-bacterial activity and systemic toxicity also.

6)Types and number of Microorganisms present:

-The efficiency of disinfections greatly depends on nature and number of contamination of
microorganisms and especially on presence or absence of bacterial spore.
-The spores are difficult to destroy but some disinfectant is destroyed the bacteria
Ex-Aldehydes, sporicidal.

7)Interfering the substance in environment:

-Materials blood, body fluids, milk, puss, food resistant / colloidal proteins may reduce the
effectiveness of disinfectants of present in small amount.
-The presence of oil and fat marketedly reduce the disinfecting ability of phenolic.

8)Potentiation, synergism & antagonism of disinfectants:

Potentiation:
-potentiation of disinfectant leads to enhance the antimicrobial activity.
Synergism:
-It is increasing in the antimicrobial activity.
Antagonism:
-This effect is often shown by two antimicrobial agents which is decrease the activity.

*Types of disinfectant:
 a) Concurant:
-Carried during the coarse of patient illness.
b) Terminal:
-Infected material after removal of patient to hospital after recovery.
c) Prophylactic:
-Pasteurization of milk / water.
-Purification by chlorination.

*Classification of disinfectants:
1. Natural.
2. Physical.
3. Chemical.

1.Natural:
-Ex- Air, sunlight.
-To prevent the growth of microorganisms.
-U rays present in sunlight also have disinfection.

2.Physical:
-Heating  Thermal methods.
-Radiation.
-Filtration.

*Legal provisions:
According to drug and cosmetics act (rule 126 schedule 0) disinfectants classified as 2 types
I. Black fluid.
II. White fluid.

I.Black fluid:
-Homogenous dark brown solution of coal tar similar acid derivatives from petroleum with /
with out hydro carbon / other phenolic compounds.

II.White fluid:
-These are white / half white emulsions consisting of coal tar.
-These are one type of phenolic compounds and its derivatives.

*Evaluations of disinfectants:
1. Tube dilution method / Agar plate method.
2. Cup plate method / Cylinder plate method.
3. Ditch plate method.
4. Gradient plate method.
5. Phenol co-efficient method / rideal walker method.

1. Tube plate method / Agar plate method:

-The chemical agents are inoculated with nutrient broth or agar medium and
inoculated with test organism.
-These tubes are incubated at 30-35oc for 2-3 days and then the result in the form of
turbidity or colonies are observed.
-The result is recorded and activity of disinfectant is compared.

2.Cup plate method:

-The nutrient agar is melted cooled at the room temperature and poured into the Petri
dish.
-Spread the 0.2ml of known concentration of inoculum on the surface of solidified
agar (spread plate method).
-Cups are cavities are made by sterile borer now 0.2ml of drug is pored into the cups
of agar plate and then incubated at 37oc for 24 hours.
-The rugs has any bacterial activity it will show the zone of inhibition.

3.Ditch plate method:

-Nutrient agar is melted cooled to the suitable poured into Petri dish.
-Solidified media is cut with sterile blade make the DITCH.
-The drug is poured very carefully into the ditch .
-Various microorganisms are streaked on sides of ditch.
-This method s used to find out the potency of the drug against the various
microorganisms by the means of inhibition of growth on streaking area.

4.Gradient plate method:

-This technique is used to isolate the resist mutant.
-The petri dish is kept in slanting position and sufficient amount of agar is poured and
solidified in slanting position.
-Another layer of agar is poured over it which contains the antibiotic solution and
solidified it.
-After solidification 0.2ml of bacterial solution culture was spread over the solid
surface.
-Incubate at 37oc for 24-48 hours.
-The microorganisms will grow where the concentration of the drug is below the
critical level.
-Antibiotic is isolate on lower layer and gradient of concentration will be produced.
-Thus the resistant nutrient can be isolated.

5.Phenol co-efficient method:

-Phenol co-efficient test is for disinfectant miscible with water and which exert their
anti-microbial action in manner similar to that of phenol.
o Test organism Salmonella Typhi.
o Standard dis-infectant Phenol.
-Different dilutions of test disinfectant and phenol are prepared and 5ml of each
dilution is inoculated with 0.5ml of broth culture of the organism for the 24 hours.
-All tubes are placed in 17.5oc water bath.
-Sub culture of each reaction mixture are taken and transferred to the 5ml of sterile
broth after 2.5, 5, 7.5, 10min.
-The broth are incubated at 37oc for 48-72 hours and are examined for presence or
absence of growth.

Time of Exposure (min)

Disinfectant Dilution
2.5min 5min 7.5min 10min

Phenol 1:100 + + - -

1:1000 - - - -

1:1100 + - - -
Unknown
substance
1:1200 + -
+ -

+ - -
1:1300 +

*Rideal walker coefficient=Dilution of unknown concentration which kill

In 7.5min but not in 5min
Dilution f which kills unknown bacteria
In 7.5min but not in 5min.
= 1200/100
= 12