Disinfectant Efficacy Study

REVIEW OF LITERATURE
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INTRODUCTION
Requirements for aseptic processing areas include readily cleanable floors, walls, and ceilings
that have smooth, non-porous surfaces; particulate, temperature, and humidity controls; and
cleaning and disinfecting procedures to produce and maintain aseptic conditions. These
conditions combined with careful and thorough evaluation of the chemical agents used for the
cleaning and disinfection program should lead to achieving the specified cleanliness
standards and control of microbial contamination of products. In recent years, the use of
disinfectants in pharmaceutical, biotechnology, medical device facilities, and associated
controlled environments has been the subject of scrutiny by regulatory agencies.
An effective cleaning and disinfection program in aseptic processing areas of a Good

Manufacturing Practice (GMP) facility is critical to assure the quality of the products.
Manufacturers are being held to a high standard when it comes to product sterility and
regulatory agencies are increasingly asking for validation data to support sanitization and
disinfection procedures. Regulatory authorities now expect evidence of the efficacy of
disinfection agents against environmental isolates. The FDA Guideline for Aseptic Processing
states, “The suitability, efficacy, and limitations of disinfecting agents and procedures should
be assessed. The effectiveness of these disinfectants and procedures should be measured by
their ability to ensure that potential contaminants are adequately removed from surfaces”.
Basic knowledge regarding the effectiveness of different chemical agents against vegetative
bacteria, fungi, and spores will aid in selecting chemical agents.
A good understanding of test methods used to assess disinfectant effectiveness is important.

Most methods are adaptable allowing the user to customize the methods to their specific
requirements.
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H I S T O RY O F D I S I N F E C TA N T S
JOSEPH LISTER & ANTISEPTIC SURGERY
Introduction: By the middle of the nineteenth century, post-operative sepsis infection

accounted for the death of almost half of the patients undergoing major surgery.
In 1839 the chemist Justin von Liebig had asserted that sepsis was a kind of combustion
caused by exposing moist body tissue to oxygen. It was therefore considered that the best
prevention was to keep air away from wounds by means of plasters, collodion or resins.
Joseph Lister (1827-1912)
Joseph Lister, a British surgeon, doubted this explanation. For many years he had explored
the inflammation of wounds, at the Glasgow infirmary. These observations had led him to
consider that infection was not due to bad air alone, and that 'wound sepsis' was a form of
decomposition.
When the Regius Professorship of Surgery at Glasgow University fell vacant in 1859 Lister
was selected from seven candidates. In August 1861 he was appointed surgeon at the
Glasgow Royal Infirmary and put in charge of its new surgical building.
The hope was that the new building would decrease the number of deaths caused by what was
then called hospital disease (now known as operative sepsis). This proved a vain hope when
Lister reported that between 45 and 50 percent of his amputation cases died from sepsis
between 1861 and 1865 in his Male Accident Ward. It was in this ward that Lister began his
experimental work with antisepsis.
Having tried methods to encourage clean healing, with little or no success, Lister began to
form theories to account for the prevalence of sepsis. He discarded the popular concept of
direct infection by bad air and postulated that sepsis might be caused by a 'pollen-like dust'.
Although, there is no evidence
GERM CONNECTION WITH WOUND SEPSIS
When, in 1865, Louis Pasteur suggested that decay was caused by living organisms in the air,
which on entering matter caused it to ferment, Lister made the connection with wound sepsis.
A meticulous researcher and surgeon, Lister recognized the relationship between Pasteur's
research and his own. He considered that microbes in the air were likely causing the
putrefaction and had to be destroyed before they entered the wound.
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In the previous year Lister had heard that 'carbolic acid' was being used to treat sewage in
Carlise, and that fields treated with the affluent were freed of a parasite causing disease in
cattle.
CARBOLIC ACID
Lister now began to clean wounds and dress them using a solution of carbolic acid.
He was able to announce at a British Medical Association meeting, in 1867, that his wards at
the Glasgow Royal Infirmary had remained clear of sepsis for nine months.
Operations using Lister’s carbolic spray invented in 1869 although his methods initially met
with indifference and hostility, doctors gradually began to support his antiseptic techniques.
In 1870, Lister’s antiseptic methods were used, by Germany, during the Fransco-Prussian war
saving many Prussian soldiers’ lives. In Germany, Robert Koch (1878) demonstrating the
usefulness of steam for sterilizing surgical instrument and dressings.
German surgeons were beginning to practice antiseptic surgery, which involved keeping
wounds free from micro-organisms by the use of sterilized instruments and materials.
Lister became aware of a paper published (in French) by the French chemist Louis Pasteur
which showed that rotting and fermentation could occur without any oxygen if micro-
organisms were present. Lister confirmed this with his own experiments. If micro-organisms
were causing gangrene, the problem was how to get rid of them. Pasteur suggested three
methods: filter, heat, or expose them to chemical solutions. The first two were inappropriate
in a human wound, so Lister experimented with the third.
Carbolic acid (phenol) had been in use as a means of deodorizing sewage, so Lister tested the
results of spraying instruments, the surgical incisions, and dressings with a solution of it.
Lister found that carbolic acid solution swabbed on wounds markedly reduced the incidence
of gangrene and subsequently published a series of articles on the Antiseptic Principle of the
Practice of Surgery describing this procedure in Volume 90, Issue 2299 of The Lancet
published on 21 September 1867.
He also made surgeons wear clean gloves and wash their hands before and after operations
with 5% carbolic acid solutions. Instruments were also washed in the same solution and
assistants sprayed the solution in the operating theatre. One of his conclusions was to stop
using porous natural materials in manufacturing the handles of medical instruments.
As the germ theory of disease became more widely accepted, it was realized that infection
could be better avoided by preventing bacteria from getting into wounds in the first place.
This led to the rise of sterile surgery. Some consider Lister "the father of modern antisepsis."
In 1879 Listerine mouthwash was named after him for his work in antisepsis. Also named in
his honor is the bacterial genus Listeria, typified by the food-borne pathogen Listeria
monocytogenes.
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The first major development towards modifying basic antimicrobial agents to make them
more specialized, more effective, was in 1906 when Bechhold and Ehrlich synthesized the
bis-phenols. They found that by linking two phenol rings directly or by mean of an alkaline
group, the bactericidal and bacteriostatic effects of the phenols were greatly enhanced.
Further work was done on the bis-phenol (During 1931) leading eventually to the synthesis of
hexachlorophene (Gump 1941; Smylie et al, 1959).
Iodine has also been modified for use as an antiseptic. The Iodine molecules can be added to
other substances which have the ability to increase the antimicrobial qualities of the elements,
whilst reducing the irritancy of iodine. The more recent modification of iodine has been the
production of iodine-alcohol tinctures and polyvinyl-pyrrolidon-iodine complexes
(Gershinfield 1957).
Alcohols, whilst proven to be excellent antimicrobial agents, have hardly been modified since
the 1880. It has been shown that, for instance, an increase in the chain length of amyl
alcohols will increase their bactericidal activity (Wirgins 1904). Alcohols can also be
incorporated into other antiseptics to boost the antimicrobial activity.
Some compounds which have been developed as antiseptics include the surface active
cationic quaternary ammonium compounds (Jacobs, 1916), although these compounds did
not have their germicidal activity realized until 1935 by Domagk. During the latter part of
19th century, many test procedures were made available for examining the efficacy of skin
antiseptic (Sykes 1985).
a) Skin infection test- which measure the antiseptic efficacy of germicides in contact
with artificially contaminated skin wounds
b) Skin penetration test- which asses the retention of activity of an antiseptic by the
skin over a period of time
c) Skin disinfection test- which evaluate antiseptic properties on intact skin.
While the scientific application of disinfectants, sterilants, and preservatives is limited

to, at most, the past 150 years, empirical practices that were more or less effective go
back to ancient times. Bible is filled with injunctions on cleanliness, strict dietary
rules, regulations regarding lepers, care regarding wastes, these matters were almost
an obsession.
The man who deserves the laurel wreath for having first recorded adequate experiment proof
as well as interpretation of the role of a microorganism in the causation of disease is Issac-
Benedict Prevost of Montauban, France. He also used chemical disinfectants and showed in
field tests that they were effective in preventing infection. In prevention of human disease,
the first half of the 19th century was a period of awakening. The association between foul
odors and disease was the basis for selection prior to the establishment of the germ throng of
human disease.
Kronig and Paul in their classical research published in 1897 laid the foundation of
scientific knowledge of chemical disinfection. They observed that disinfectants can be
accurately compared only when tested under controlled conditions. The number of bacteria
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must be constant and brought in to the contact with the disinfectant without the interference
of other organic matter such as the nutritive medium. The action of the disinfectant must be
promptly arrested after a stated period of time and the bacteria transferred to the most
favorable medium at optimum temperature. The results of test are determined by accurate
count of survivors in plate cultures.
Cleaning methods today in hospitals have largely been developed from studies of dust control
carried out in the early 1960s. These studies examined the efficacy of different cleaning
methods, by comparing not only various techniques for removing dust, but also numerous
disinfectants used for the chemicals treatment of environmental contamination.
From ‘cleaning’ point of view, the pharmaceutical units may be regarded as consisting of two
distinct types of environment; ‘sterile’ production areas where injections, vaccine’s and other
products are prepared from living microorganisms and ‘non-sterile’ production areas where
medicines for internal and external use are made in bulk. Difference between the two types
environment have become more marked in recent years, largely as a result of the introduction
of clean room technology to sterile production area, microorganism may be introduced to the
environment via the raw materials, air and water supplies, packing components, processing or
cleaning equipment; or by the staff who work in the unit. Aerobic spore bearers are generally
accepted as being inevitable contaminants in many supplies, and hence are often found in
pharmaceutical units. Likewise, unless protective clothing is worn by the staff, large numbers
of Gram positive cocci may be shed from the skin into the manufacturing environment. It is
known that contaminated medicines may serve as a vehicle for the spread of cross-infection
when used in the wards. In particular, certain disinfectants are susceptible to contamination
by Pseudomonas aeruginosa, when incorrectly made or used and ironically made or used and
ironically these may contributed to, rather than reduce the microbial flora of the area to be
cleaned.
Studies in pharmaceutical units have shown that floors and other horizontal surfaces are often
heavily contaminated with microorganisms. Washing these surfaces with hot water and soap
alone removes a large proportion of contaminants; washing with disinfectants can result in a
significant improvement. Surfaces such as walls and ceilings generally have a much smaller
microbial population if kept in good repair and hence requires cleaning much less often than
floors. Several studies have suggested that surfaces cleaned too vigorously and frequently
may become thereby encouraging environmental contamination.
The number of range of microorganisms that are resistant to one or more antibiotic is a
continual and increasing problem, especially in hospitals. With the emergency of pathogens
such as methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin resistant
enterococci (VRF) and gram-negative bacteria, for example, Pseudomonas aeruginosa and
Acinetobacter species which are resistant to multiple antibiotics, there is an increasing need
for effective disinfection. This is especially true in clean rooms where pharmaceuticals,
dressing and instrument are prepared and sterilized.
In general, there were no disinfection policies in place for the use and concentration of the
disinfectants available since last many years. The situation has changed markedly and now
many pharmaceutical industries have such policies, but implements are still frequently
unsatisfactory.
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Clearly, there is a need for pharmaceuticals unit to understand the role and uses of
disinfectants, the factor that influences their activity, the possibility that microbial resistance
can arise and the relevance, if any of rotation of disinfectants.
Disinfectant: an agent that frees from infection; usually, a chemical agent which destroys
disease germs or other harmful microorganisms; commonly used as substances applied to
inanimate objects. While this definition has been in general use for many years, it is
sometimes misused, as in “skin disinfectant”; “skin antiseptic” is more appropriate in
describing a germicide for use on the body or inanimate surfaces.
One another definition employed for disinfectant is- an agent that frees from infection;
usually a chemical agent which destroys disease germs or other microorganisms or
inactivates viruses. The official definition of the word “Disinfection” adopted by the
American public health Association (1950) is as follows: “Disinfection – killing of
pathogenic agents by chemical or physical means directly applied”.
A sound cleaning and sanitization program is needed for controlled environment used in the
manufacture of pharmacopeial articles to prevent the microbial contamination of these
products. Sterile drug products may be contaminated via their pharmaceutical ingredients,
process water, packaging components, manufacturing environment processing equipment and
manufacturing operators. Current good manufacturing practices (cGMP) emphasize the size,
design, construction and location of buildings and construction materials and the appropriate
material flow to facilitate cleaning, maintenance and proper operations for the manufacture of
drug products. When disinfectants are used in a manufacturing environment, care should be
taken to prevent the drug product from becoming contaminated form chemical disinfectant as
a result of the inherent toxicity of the disinfectants. The requirement for aseptic processing
include readily cleanable floors, walls and ceilings that have smooth and non porous surfaces;
particulate, temperature and humidity controls; cleaning and disinfecting procedures to
produce and maintain aseptic conditions. The cleaning and sanitization program should
achieve cleanliness standards, control microbial contamination of products, and be designed
to prevent chemical contamination of pharmaceutical ingredients; products contact surfaces
or equipments, packaging materials and ultimately the drug products. These principles also
apply to non sterile dosage forms where the microbial contamination is controlled by the
selection of appropriate pharmaceutical ingredients, utilities, manufacturing environments,
sound equipments cleaning procedures, products especially formulated to control water
activity, inclusion of suitable preservatives, and products packaging design.
In addition to disinfectants, antiseptics are used to decontaminate human skin and exposed
tissues and may be used by personnel prior to entering the manufacturing area. Chemical
sterilants mat be used to decontaminate surfaces in manufacturing and sterility testing areas.
Furthermore, sterilants may be used for the sterilization of pharmacopeial articles, and UV
irradiation may be used as a surface sanitizer.
This general introduction will discuss the selection of suitable chemical disinfectants and
antiseptics:
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• The demonstration of their bactericidal, fungicidal and sporicidal efficacy;
• The application of disinfectant in the sterile pharmaceutical manufacturing

area;
• Regulations and safety consideration.
Definitions:-
Antiseptics: An agent that inhibits or destroys microorganisms on living tissue including
skin, oral cavities and open wounds.
Chemical disinfectants: A chemical agent used on inanimate surfaces and objects to destroy
infectious fungi, viruses and bacteria but not necessarily their spores. Sporicidal and antiviral
agents may be considered a special class of disinfectants. Disinfectants are often categorized
as high level, intermediate level, and low level by medically oriented groups based upon their
efficacy against various microorganisms.
Decontamination: The removal of microorganisms by disinfection or sterilization.
Disinfectants: A chemical or physical agent that destroys or removes vegetative forms of
harmful microorganisms when applied to a surface.
Sanitizing agents: An agent which reduce microbes on inanimate surfaces, the number of all
forms of microbial life including fungi, viruses and bacteria.
Sporicidal agents: An agent that destroys bacterial and fungal spores when used in sufficient
concentration for a specified contact time. It is expected to kill all vegetative microorganisms.
Sterilants: An agent that destroys all forms of microbial life including fungi, viruses and all
forms of bacteria and their spores. Sterilants are liquid or vapor-phase agents.
Microorganisms differ greatly in their resistance to disinfecting agents. The order of

resistance of clinically significant microorganisms to chemical disinfectants from most to
least resistant.
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Table-1. The resistance of some clinically important microorganisms to chemical
disinfectants (Listed in order of decreasing resistance).
1) Type of Microorganisms Examples
1) Bacterial spores Bacillus subtilis and Clostridium sporogenes.
2) Mycobacteria Mycobacterium tuberculosis.
3) Non lipid-coated viruses Poliovirus and Rhinovirus.
4) Fungal spores and vegetative molds Trichophyton, Crytococcus and Candida sp.and
yeasts.
5) Vegetative bacteria Pseudomonas.aeruginosa,

Staphylococcus aureus and
Salmonella etc.
6) Lipid-coated viruses Herpes simplex virus, Hepatitis B virus, HIV
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C L A S S I F I C AT I O N O F D I S I N F E C TA N T S
Chemical disinfectant is classified by their chemical type. This includes Aldehydes, alcohol,
halogens, and per-oxides, quaternary ammonium compounds and phenolic compounds.
Table-2: General classification of Antiseptics, Disinfectants and Sporicidal Agents.
Chemical Entity Classification Examples

1) Aldehydes Sporicidal agents 2% Gluteraldehyde
2) Alcohol General purpose disinfectants, antiseptic, 70% Isopropyl

antiviral agents. alcohol
70% Alcohol.
3) Chlorine and Sporicidal agents 0.5% Sod.
Sod.hypochlorite hypochlorite
4) Phenolics General purpose disinfectant 500µg per gm chloro-
xylenol.
5) Ozone Sporicidal agent 8% gas by weight
6) H2O2 Vapor phase sterilant, liquid sporicidal 4µg per gm H2O2
agent, antiseptic vapor
10-25% solution, 3%
solution.
7) Substituted Antiseptic agents 0.5% Chlorhexidine
Diguanides gluconate.
8) Per-acetic acid Liquid sterilant, vapor phase sterilant 0.2% Per-acetic acid,
1µg per gm per-acetic
acid.
9) Ethylene oxide Vapor-phase sterilnt 660µg per gm
ethylene oxide.
10) Quaternary General purpose disinfectant, antiseptic 200µg per gm
Amm. Benzalkonium
compounds chloride
11) β- Propiolactonic Sporicidal agent 100µg per gm β-
Propiolactone.
Under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA), the Environment
protection agency (EPA) registers chemical disinfectant marketed in the United States and
requires manufacturers to supply product information on the use dilution, and type of
microorganisms killed and the necessary contact time. Certain liquid chemicals sterilizers
intended for use on critical or semi critical medical device are defined and regulated by the
U.S Food and Drug Administration (FDA).Selection of an antiseptic for hand and surgical
site disinfection:
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SELECTION OF AN ANTISEPTIC FOR HAND AND SURGICAL SITE
DISINFECTION:
Hands and surgical sites are disinfected in a hospital setting to reduce the resident flora and to
remove transient flora (e.g. – Streptococcus pyogenes and methicillin resistant
Staphylococcus aureus and Pseudomonas aeruginosa) that have been implicated in
hospital- associated infection. Use of antiseptic to disinfectant hands has been shown to be
more effective than soap and water in reducing the counts of bacteria on the skin; repeated
antiseptic use further reduces these counts. These principles may be applied to clean room
operation in the pharmaceutical industry.
MECHANISM OF DISINFECTANT ACTIVITY
Target Disinfectants
Cell wall Formaldehyde, hypochlorite’s and mercurials

1) Cytoplasmic membrane, action on Anilides and hexachlorophene
membrane potential
2) Membrane enzymes, action on Hexachlorophene
electron transport chain
3) Action on ATP Chlorhexidine and ethylene oxide
4) Action on enzymes with –SH group Ethylene-oxide, Gluteraldehyde, Hydrogen
peroxide, hypochlorite’s, Iodine and
mercurial’s
5) Action on general membrane Alcohols, Chlorhexidine and quaternary
permeability ammonium compounds.
6) Cell contents, general coagulation Chlorhexidine, Aldehydes, Hexachlorphene
and quaternary ammonium compounds
7) Ribosome’s Hydrogen peroxide and mercurial’s
8) Nucleic acids Hypochlorite’s
9) Thiol-groups Ethylene oxide, gluteraldehyde, hydrogen
peroxide, hypochlorite’s and mercurial’s
10) Amino groups Ethylene oxide, gluteraldehyde and
hypochlorite’s
11) General oxidation Ethylene oxide gluteraldehyde and
hypochlorite’s
MICROBIAL RESISTANCE TO DISINFECTANTS:
The development of microbial resistance to antibiotics is a well described phenomenon. The

development of microbial resistance to disinfectant is less likely, as disinfectants are more
powerful biocidal against low population of microorganisms usually not growing actively, so
the selective pressure for the development of resistance is less profound. However, the most
frequently isolated microorganism from an environmental monitoring program may be
periodically subjected to use dilution testing with the agents used in the disinfection program
to confirm their susceptibility.
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DISINFECTANT CHALLENGE TESTING
The EPA requires companies that register public health antimicrobial pesticide products
including Disinfectants, Sanitization agents, Sporicidal agents and sterilants to ensure the
safety and effectiveness of their products before they are sold or distributed. Companies
registering those products must address the chemical composition of their product include
toxicology data to document that their product is safe? If used as directed on the label,
include efficacy data to document their claims of effectiveness against specific organisms and
to support the directions for use provided in the labeling and provide labeling that reflects the
required elements for safe and effective use. While these directions provide valuable
information, they may not be helpful in terms of the products use as disinfectants in a
manufacturing environment.
To demonstrate the efficacy of a disinfectant with a pharmaceutical manufacturing
environment, it may be deemed necessary to conduct the following tests;
1) In Use-dilution method: - Screening disinfectant for their efficacy at various
concentration and contact times against a wide range of standard test organisms and
environment isolates.
2) Surface challenge test: - Using standard test micro-organisms and test

microorganisms that are typical environmental isolates, applying disinfectant to
surfaces at the selected used concentration with specified contact time, and
determining the log reduction of the challenge microorganisms.
3) A statistical comparison of the frequency of isolation and numbers of microorganisms

isolated prior to and after the implementation of a new disinfectant. This is considered
necessary because critical process steps like disinfection of aseptic processing areas as
required by EMP regulations need to be validated, and the EPA registration
requirements do not address how disinfectants are used in the pharmaceutical,
biotechnology, and medical device industries.
For the surface challenge tests, the test organisms are enumerated using swabs, surface
rinse, or contact plate methods. Neutralizers that inactivate the disinfectant should be
included in either the diluents or microbiological media used for microbial enumeration or
both.
TABLE: - NEUTRALIZING AGENTS FOR COMMON DISINFECTANT
Disinfectant Neutralizing Agents
1) Alcohols Dilution or polysorbate 80

2) Gluteraldhyde Glycine and Sodium bisulfate
3) Sodium hypochlorite’s Sodium thiosulfate
4) Chlorhexidine Poltsorbate 80 and lecithins
5) Mercuric chloride Thioglycolic acid
6) Quaternaryammonium Polysorbate 80 and lecithins
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compounds
Universal neutralizer’s broths may be formulated to contain a range of neutralizing agents.

For example, Dey-Engley (D/E) broth contains 0.5% polysorbate 80, 0.7% lecithin, 0.1%
Sodium thioglycolate, 0.6% Sodium thiosulfate, 0.25% Sodium biosulfate, 0.5% tryptone,
0.25% yeast extract, and 1.0% dextrose; letheen broth contains 0.5% polysorbate 80, 0.07 %
lecithin, 1.0% peptamin, 0.5% beef extract, and 0.5% Sodium chloride; and Tryptone-
Azolecithin-Tween (TAT) broth base contains 4.0%(v/v) polysorbate 20, 0.5% lecithin, and
2.0% Tryptone.
In practice, sufficient organisms need to be inoculated on a 2 inch× 2 inch square of the
surface being decontaminated, i.e. a coupon, to demonstrate at least a 2 (bacterial spore) to 3
(for vegetative bacteria) log reduction during a predetermined contact time ( i.e. 10 minutes
over and above the recovery observed with a control disinfectant application). The efficacy of
the neutralizers and their ability to recover inoculated microorganisms from the material
should be demonstrated during the use dilution or surface challenge studies. Points to be
remember are that disinfectants are less effective against the higher numbers of
microorganisms used in laboratory challenge tests than they are against the numbers that are
found in clean rooms; that inoculate from the log growth phase that are typically employed in
laboratory tests are more resistant, with the exception of spores formed during the static
phase, than those from a static or dying culture or stressed organisms in the environment; and
that microorganisms may be physically removed during actual disinfectant application in the
manufacturing area.
Although not all inclusive, typical challenge organisms that may be employed are listed in
Table;
TYPICAL CHALLENGE ORGANISMS
AOAC Challenge Organisms Isolates Typical Environmental
Bactericide: Escherchia. coli Bactericide: Micrococcus. luteus
Staphylococcus. aureus Streptococcus. Epidermis
Pseudomonas aeruginosa Coynebacteria.jeikeium
Fungicide: Penicillium. Chrysogenum Fungicide: C.albicans,

Aspergillus niger
Penicillium. Chrysogenum
Sporicide: Bacillus. Subtilis Sporicide: Bacillus. Sphaericus
Bacillus.thuriengiensis
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Because a wide range of different materials of construction are used in clean rooms and other
controlled areas, each material needs to be evaluated separately to validate the efficacy of a
given disinfectant.
Table contains a list of common materials used in clean room construction.
Table: - Typical surfaces to be decontaminated by disinfectants in a Pharmaceutical
manufacturing Area.
Materials Application
1) Stainless steel Work surfaces, filling equipments and tanks.
2) Glass Windows and vessels.
3) Plastic, Vinyl Curtains.
4) Plastic, Polycarbonate Insulation coating.
5) Lexan(Plexiglas’s) Shields.
6) Epoxy-Coated gypsum Walls and Ceilings.
7) Fiber glass-Reinforced plastic Wall paneling.
8) Terrazzo tiles Floors.
DISINFECTION IN CLEANING AND S A N I T I Z AT I O N

PROGRAM
The selection of suitable disinfectants and the verification of their effectiveness in surface
challenge testing are critical in the development of a cleaning and sanitization program. Issue
associated with the successful implementation of such a program are the development of
written procedure, staff training, decisions on disinfectant rotation, institution of application
methods and contact times, environmental monitoring to demonstrate efficacy, and personnel
safety. Staff involved in disinfection requires training in microbiology, industry practices for
cleaning and sanitization, safe handling of concentrated disinfectants, the preparation and
disposal of disinfectant, and appropriate application methods.
It should be emphasized that the preparation of the correct dilution is critical because many
disinfectant failures can be attributed to use of disinfectant solution that are too dilute.
Typically disinfectants used in aseptic processing and filling areas are diluted with sterile
purified water, and are prepared aseptically. Alternately, the disinfectant may be diluted with
purified water, and then sterile filtered to eliminate microorganisms that may potentially
persist in a disinfectant. Dilute disinfectants must have an assigned expiration dating justified
by effectiveness studies.
Because it is theoretically possible that the selective pressure of the continuous use of a single
disinfectant could result in the presence of disinfectant could result in the presence of
disinfectant- resistant microorganisms in a manufacturing area, in some quarters the rotation
of disinfectant has been advocated. However, the literature supports the belief that the
exposure of low numbers of microorganism on facility equipment surfaces within a clean
room where they are not actively proliferating will not result in the selective pressure that
may be seen with the antibiotics. It is prudent to augment the daily use of a bactericidal
disinfectant with weekly (or monthly) use of a sporicidal agents is not generally favored
because of their tendency to corrode equipment and because of the potential safety issues
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with chronic operator exposure. Other disinfection rotation schemes may be supported on the
basis of a review of the historical environment monitoring data. Disinfectants applied on
potential product contact surfaces are typically removed with 70% alcohol wipes. The
removal of residual disinfectants should be monitored for effectiveness as a precaution
against the possibility of product contamination
The greatest safety concerns are in the handling of concentrated disinfectants and the mixing
of incompatible disinfectants. For example, concentrated sodium hypochlorite solutions (at a
concentration of more than 5%) are strong oxidants and will decompose on heating, on
contact with acids, and under the influence of light, producing toxic and under the corrosive
gases including chlorine. In contrast, dilute solutions (at a concentration of less than 0.5%)
are not considered as hazardous. Under no circumstances should disinfectants of different
concentration be mixed. Material safety data sheets for all the disinfectants used in a
manufacturing area should be available to personnel handling the disinfectant preparation,
and personnel must be trained in the proper use of this equipment. Safety showers and eye
wash stations must be situated in the work area where disinfectant solutions are prepared.
APPLICATIONS OF DISINFECTANTS
Disinfectant Uses
Alcohols Ethyl or isopropyl alcohol at 70-80% concentration is a good
general purpose disinfectant; not effective against bacterial spores.
Phenols Effective against vegetative bacteria, fungi and viruses containing
liquids, unpleasant odor.
Formaldehyde Concentration of 5-8% formalin is a good disinfectant against
vegetative bacteria, spores and viruses; known carcinogen;
irritating odor.
Quaternary Ammonium Cationic detergents are strongly surfaced active; extremely
Compounds effective against lipoviruses; ineffective against bacterial spores;
may be neutralized by anionic detergents (i.e. soaps).
Chlorine Low concentrations (50-500 ppm) are active against vegetative
bacteria and most viruses; higher concentrations (2,500 ppm) are
required for bacterial spores; corrosive to metal surfaces; must be
prepared fresh; laundry bleach (5.25% chlorine) may be diluted
and used as a disinfectant.
Recommended for general use; effective against vegetative
Iodine bacteria and viruses; less effective against bacterial spores;
Wescodyne diluted 1 to 10 is a popular disinfectant for washing
hands.
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FACTORS AFFECTING DISINFECTION
The following are six primary variables that influence the efficacy of disinfection:
1) Nature of the item to be disinfected:
The rougher the surface, the longer the contact time required for disinfection.
2) Number of microorganisms present:
The number of microorganism present will lengthen the time for effective disinfection to take
place. In general, higher numbers of organisms require more time for disinfection.
3) Resistance of microorganisms:
Some microorganisms are more resistant to disinfection than others. The generally accepted
order from the most resistance to the least resistant is: bacterial spore, mycobacteria,
hydropjillic viruses, fungi, vegetative bacteria, lipid viruses. Disinfecting a spill with a small
concentration of bacterial spores will require longer disinfection time than a large concentration
of lipid viruses.
4) Type and concentration of disinfectant used:
Resistance of microorganisms depends on the type of disinfectant used. A particular

microorganism may be more resistant to one type of disinfectant than another. For instance,
alcohol (Isopropyl or ethyl) is effective against vegetative bacteria and most lipophilic viruses.
Many disinfectants are broad spectrum; that is, effective against all or most forms of microbial
life. Some non-broad spectrum disinfectants include phenolics and quaternary ammonium
compounds. Some broad spectrum disinfectants include Gluteraldehydes, sodium
hypochlorites, and hydrogen peroxides. Alcohols lie somewhere in between these two.
The concentration of a particular disinfectant effects disinfection. In most cases, a higher

concentration increases microbial killing power and decreases time necessary for disinfection.
However, some disinfectants are not as effective in higher concentrations. Iodophors must be
diluted according to the directions on the label; over-diluting or under-diluting may
substantially lower the microbial potency. Alcohols used in concentration above 90% are less
effective because the water added to dilute the alcohol allows it to penetrate better and reach its
target. Optimal concentration range is between 70-90%.
16
5) Presence of organic material:
The presences of organic soiling matter will compromises disinfection. Blood, blood product,
bodily fluids and feces contain significant amounts of proteins, and protein will bind and
inactivate some disinfectants or slow their action. Therefore, in the presence of large amounts
of protein, a higher concentration of disinfectant and longer contact time will be necessary to
achieve maximal disinfection.
17
6) Duration of exposure and temperature:
Duration of exposure and temperature influences the disinfection process. The longer
the duration of exposure, the higher the degree of disinfection achieved. Some
disinfectants require a longer contact time to achieve killing, and some
microorganisms need longer exposures to be killed. Higher temperature increases the
killing power of most disinfectants, where lower temperature may slow the killing
power of most disinfectants. .
The following disinfectants were taken up for the Efficacy and Shelf life study:
General Description of disinfectants used:
1) Incidur:
Incidur is a broad spectrum; Microbiocidal action including HIV, Hepatitis B etc.It

shows long residual effect with sustained release. It is manufactured by Henkel Germany.
It is purposely used for surface disinfection and fumigation.
Active ingredients in 100 gms solutions:
• Glyoxal…………..8.8gm.
• Gluteral…………..4.5gm.
Range of application:
 High risk areas like Operation theatres, Intensive care Unit (ICU), laboratories and
drug manufacturing units.
 Commonly used for general purpose in laboratories and pharmaceuticals units.
Mode of Action:
 It interferes with enzymes with –SH groups and Thiol group of proteins.
 It is also responsible for general oxidation of cellular components in microbial cells.
Recommended concentration and contact time (According to manufacturer):
• According to manufacturer (Henkel Germany), Incidur is effective at 1%

concentration for 1 hour exposure or contact time.
2) Levermed:
Levermed is broad spectrum hand disinfectant. It acts as Bactericidal, Sporicidal,

Fungicidal and Virucidal agent. It is manufactured by Universal Healthcare Silvassa.
18
It is marketed by Johnson Diversy. It is purposely used for hand disinfection and
laboratory practices. Levermed is ready to use prepared hand disinfectant.
Active Ingredients (w/w):
• n- popanol………………………50%
• Iso-propyl Alcohol...……………20%
• Benzalkonium chloride………...0.5%
• Emollents.
Range of Application:
• Industrial Quality Control Laboratories, Research Institutes, Academics, Drug

Manufacturing units.
Mode of Action:
• It generally shows action towards membrane permeability. It coagulates cellular

components.
Recommended concentration and contact time (According to manufacturer):
• According to manufacturer (Universal Healthcare, Silvassa), Levermed is effective at

10 minutes exposure or contact time.
3) Oxivir:
Oxivir is broad spectrum environmental cleaner & disinfectant. It is oxygen based

Bactericidal, Sporicidal, Virucidal, Fungicidal, Yeasticidal. Oxivir is concentrated
oxygen based liquid cleaner disinfectant for cleaning and disinfectant of all water
resistant surfaces. It is manufactured by Johnson Diversy. It is effective against
Bacteria, Fungi, Yeasts, Virus. It is purposely used for Fumigation, Cleaning and
disinfecting equipment. It is compatible with metals and surfaces used in hospitals.
However, material compatibility on a small and inconspicuous place is recommended
before use.
Composition:
Hydrogen peroxide
Alkyl Benzene sulphonic acid
19
Alcohol ethoxylate (C12-15) 3EO
Hydroxyethene-1-1-diphosphonic acid
2- butoxyethanol
Feature Advantage Benefits
Accelarated hydrogen Higher grade disinfectant Broad spectrum Microbial kill

peroxide
Aldehyde free product No negative occupational Eco-friendly and user friendly

hazards
Cleaner and Single shot product Time and cost effective

disinfectant
 Oxivir is purposely used in Hospitals, laboratories, Institutes and Drug manufacturing

units.
Recommended Concentration and contact time:
 According to manufacturer (Johnson Diversy), Oxivir is effective at 2.5% for 5

minute contact time.
Mode of Action:
 Oxivir shows action on enzymes with –SH group.
 Oxivir also cause general oxidation of cellular components.
4) Virex II 256:
Virex II 256 is high level instrument & surface disinfectant Bactericidal, Sporicidal,
Virucidal, Fungicidal, Yeasticidal. Virex II 256 is a high level disinfectant of surfaces
and critical instruments (which come in contact with blood stream or normally sterile
areas of the body, such as rigid and flexible surgical instruments) in healthcare
application. . It is purposely used for Cleaning and disinfecting equipment. It is
compatible with metals and surfaces used in hospitals. However, material
compatibility on a small and inconspicuous place is recommended before use. Virex II
256 is blue colored liquid disinfectant manufactured by Johnson Diversy. It is
effective against Bacteria, Fungi, Yeasts, Virus.
Composition:
20
 Didecyl dimethyl ammonium chloride……………………..8.7%
 n-alkyl dimethyl benzyl ammonium chloride………………8.19%
 Lauryl amine amine oxide
 Ethylene diamine tetra acetic acid sodium salt
 Sodium biocarbonate
Feature Advantage Benefits
N-alkyl dimethyl benzyl Combination of most potent Broad spectrum kill

ammonium chloride & Didecyl QAC’s
dimethyl ammonium chloride
Aldehyde free product No negative occupational hazard Eco friendly and user
friendly
Cleaner and disinfectant Single shot product Time and cost effective
 Virex II 256 is purposely used for instruments disinfection. It is also used for surface
disinfection. It is also used for washing and other tiled surfaces. It is compatible with
most metals and surfaces used in pharmaceuticals units, Laboratories and Research
Institute.
Recommended Concentration and contact time:
 According to manufacturer (Johnson Diversy), Virex II 256 is effective at 0.4% for 10

minutes contact time or exposure.
Mode of Action:
 It causes general cell components coagulations.
21
AIMS AND OBJECTIVES
AIM: Assessment of efficacy and shelf life of various disinfectants used in Vaccine
formulations Plant by In-Use Dilution Method.
OBJECTIVES
• To check the efficacy of various disinfectants under laboratory conditions, stimulating

their possible practical use.
 Levermed.
 Incidur.
 Oxivir.
 Virex II 256.
22
M AT E R I A L S A N D M E T H O D O LO GY
23
TESTING THE EFFICACY OF DISINFECTANTS
Materials and Reagents:-
1) Disinfectants.
2) Microbial Cultures.
a) Staphylococcus aureus NCIM 2079.
b) Pseudomonas aeruginosa NCIM 2200.
c) Bacillus subtilis NCIM 2063.
d) Escherchia coli NCIM 2065.
e) Candida albicans NCIM 3471.
f) Salmonella abony NCIM 2257.
g) Environmental Isolates of E. coli.
h) Environmental Isolates of Bacillus.
i) Environmental Isolates of Staphylococcus aureus.
Glassware and other materials:-
1) Sterile bottles.
2) Sterile test tubes.
3) Sterile Petri plates.
4) Stopwatch.
5) Test tube stand.
24
6) Inoculating loop.
7) Glass beads.
8) Micro pipette (1000µL).
9) Micro pipette (200µL).
10)Micro pipette tips (1000µL).
11)Micro pipette tips (200µL).
12) 10 mL sterile syringe.
13)0.2µm filters.
14) Anhydrous calcium chloride.
15) Anhydrous magnesium chloride.
16) 10 % dextrose solution.
17) Nutrient broth.
18)Soybean Casein Digest Agar.
19)Dey- Engley neutralizing Broth.
Preparation of solutions:
• Preparation of sterile hard water:
• Dissolve 0.304gm Calcium chloride and 0.065gm anhydrous magnesium chloride in

purified water.
• Make it up to 1 liter.
• Dispense into glass container and sterilize by autoclaving at 121°c for 30 minutes.
3) Preparation of 10 % dextrose solution:-
• Took 10gm of dextrose in a volumetric flask
• Add distill water to make up to 100 mL shake well to dissolve completely.
25
• Transfer dextrose solution in a clean glass bottle and sterilize by autoclaving at 121ºc
for 30 minutes.
3) Preparation of Nutrient broth solution:-
4) Dissolve 13 gm nutrient broth in 1000 ml water
5) Sterilize by autoclaving at 121ºc for 30 minutes.
6) To each liter of nutrient broth solution add 10 ml sterile dextrose solution for the
growth of the organism.
4) Preparation of Disinfectant solution:-
• Prepare disinfectant solution as recommendation by manufacturer.
• After dilution filter the disinfectant solution from 0.2µm filter with the help of sterile
10 ml syringe.
Procedure:-
1) Preparation of inoculums:-
 Incubate the contents of a freeze dried culture suspended in nutrient broth overnight at
37 ºc ±1 ºc for bacteria and 22.5 ºc ± 2.5 ºc for fungus.
 Inoculate the incubated culture onto nutrient agar slopes. These slants can be stored
up to 4 months at 2-8 ºc.
 Subculture from agar slopes in to 10 mL of growth medium. Incubate at 37 ± 1ºc for
± 2hr for bacteria and 22.5 ºc ± 2.5 ºc for 120 hrs for fungus.
 Subculture from the medium into fresh medium, using an inoculating loop. Inoculate
at 37 ºc ± 1 ºc for 24 ± 2 hr for bacteria and 22.5 ºc ± 2.5 ºc for 120 hrs for fungus.
 Repeat the above step daily. For the test procedure use only those cultures which have
been sub culture for at least 5 times and not more than 14 times.
 Centrifuge all cell culture until cells are compact, and remove the supernatant with a
sterile pipette.
 Re-suspend the test organism in 10 mL of growth medium and shake well for 1
minute with few sterile glass beads.
26
 Enumerate the inoculums before testing using 10 fold dilutions and pour
plate technique on SCDA plate at 30-35˚C for 48 hrs for bacteria and 22.5
± 2.5˚C for 120 hrs for fungus.
Acceptance criteria:-
The number subsequently counted must represents not less than 2×108 or not more than 2×109
organisms per ml. Retain 1 dilutions as control.
Test Procedure:-
 Add 3 mL diluted disinfectant solution to a capped sterile test tube.
 Start a timing device, immediately inoculate disinfectant solution with 1 mL of culture

and mix by swirling.
 After appropriate contact time (as recommended by manufacturer), subculture 1 drop

into each of 5 tubes containing Dey-Engley broth.
 To ensure delivery of 0.02 mL into first tube of Dey- Engley broth at exactly time, it
will be necessary to withdraw a suitable amount from the disinfectant test mixture
shortly before hand. This must be immediately preceded by vortexing.
 Mix the contents of all tubes of Dey- Engley broth by vortexing. Inoculate at 37 ºc ±
1ºc for 48 hrs for bacteria and 22.5 ºc ± 2.5 ºc for 120 hrs for fungus.
 To observe the recovery of microorganisms, after incubation take 1mL from each
Dey-Engley broth tubes, add into sterile petri plates and pour 20 mL SCDA (45 ºc) in
to each plate.
 Mix well the contents of plate. Incubate at 37 ± 1 ºc for 48 hrs for bacteria and 22.5
±2.5 ºc for 120 hrs for fungus.
Acceptance Criteria: The test passes if there is no apparent growth in at least three out of
five Dey-Engley broth tubes.
Testing of control:-
1) Dey-Engley broth contamination:-
Incubate one un-inoculated tube of Dey-Engley broth at 37 ± 1ºc for 48 hrs for
bacteria and 22.5 ºc ± 2.5 ºc for 120 hrs for fungus and examine for growth. No
growth should occur.
2) Disinfectant contamination:-
27
To 1 tube of Dey-Engley broth, add 0.02mL of disinfectant solution. Inoculate at 37 ±
1ºc for 48 hrs for bacteria and 22.5 ± 2.5 ºc for fungus and examine for growth. If
growth occurs, the test is considered invalid.
3) Fertility test:-
To 1 tube of Dey-Engley broth, add 1.0 mL of 107 dilution of culture and incubate at
37 ±1 ºc for 48 hrs for bacteria and 22.5 ± 2.5 ºc for 120 hrs for fungus and examine
for growth. If no growth occurs, the test is considered invalid.
4) In activator efficacy:-
To 1 tube of Dey-Engley broth, add 0.02 mL of disinfectant solution and 1.0 mL of

107 dilution of the culture at 37 ± 1ºc for 48 hrs for growth. If no growth occurs, the
test is considered invalid.
Purpose of Using Dey-Engley neutralizing Broth: Dey-Engley neutralizing broth is a

universal neutralizer broth. Dey-Engley (D/E) broth contains 0.5% polysorbate 80, 0.7%
lecithin, 0.1% Sodium thioglycolate, 0.6% Sodium thiosulfate, 0.25% Sodium biosulfate,
0.5% tryptone, 0.25% yeast extract, and 1.0% dextrose; letheen broth contains 0.5%
polysorbate 80, 0.07 % lecithin, 1.0% peptamin, 0.5% beef extract, and 0.5% Sodium
chloride; and Tryptone-Azolecithin-Tween (TAT) broth base contains 4.0%(v/v) polysorbate
20, 0.5% lecithin, and 2.0% Tryptone.
These ingredients are acts as a universal in activator for the inhibitory compounds. So Dey-
Engley broth is used to recover microbial inoculum after disinfectant exposure.
28
RESULTS
D I S I N F E C TA N T S E F F I C ACY
R E S U LT S
29
30
OBSERVATIONS
Disinfectant Incidur.
Manufacturer Universal Healthcare Silvasa.
Manufacturing Date March 2008.
Expiry Date Feb 2010.
Batch no 803
Recommended contact time 60 minutes.
Concentration 1% v/v
31
Disinfectant Microbial culture Contact time Dey-Engley broth tubes
(minutes)
Tube 1 Tube 2 Tube 3 Tube 4 Tube 5
Incidur S.aureus 30 minutes √ √ √ √ √
60minutes × × × × √
90 minutes × × × × ×
P.aeruginosa 30 minutes √ √ √ √ √
60 minutes √ √ √ × ×
90 minutes √ √ √ × ×
Bacillus subtilis 30 minutes √ √ √ √ √
60 minutes × × × × √
90 minutes × × × × ×
Escherchia coli 30 minutes √ √ √ √ √
60 minutes √ √ √ √ √
90 minutes √ √ √ √ √
Candida albicans 30 minutes √ √ √ √ √
60 minutes √ √ √ × ×
90 minutes √ √ √ × ×
Salmonella abony 30 minutes √ √ √ √ √
60 minutes √ √ √ √ ×
90 minutes √ √ × × ×
E. I E. coli 30 minutes √ √ √ √ √
60 minutes √ √ √ × ×
90 minutes √ √ × × ×
E.I B. subtilis 30 minutes √ √ √ √ √
60 minutes √ √ √ √ √
90 minutes √ √ √ √ √
E. I S. aureus 30 minutes √ √ √ √ √
60 minutes √ √ √ √ √
90 minutes √ √ √ √ √
32
Sign: √ = Growth.
× = No growth.
Test Controls:-
S.NO Control
1) Dey- Engley Contamination ×
2) Disinfectant Contamination ×
3) Inactivator Control √
4) Fertility Control √
Sign: √ = Growth.
× = No growth.
Enumeration of Microbial Inoculums on SCDA:-

S.No Microbial culture Cfu/mL
106 107 108 109 1010
1) Staphylococcus aureus 120 20 2 × ×
2) Pseudomonas aeruginosa TNTC 75 2 × ×
3) Bacillus subtilis TNTC 67 3 × ×
4) Escherchia coli TNTC 110 2 × ×
5) Cabdida albicans TNTC 65 3 × ×
6) Salmonella abony 140 16 2 × ×
7) Environmental isolates of Escherchia coli TNTC 15 3 × ×
8) Environmental isolates of Bacillus subtilis TNTC 25 3 × ×
9) Environmental isolates of Staphylococcus TNTC 26 2 × ×
33
aureus
Recovery:-
S.NO Microbial culture Contact Cfu/mL(Dey –Engley tube)
time
Tube 1 Tube 2 Tube 3 Tube 4 Tube
5
1) Staphylococcus aureus 30 minutes TNTC TNTC TNTC TNTC TNTC
60 minutes TNTC TNTC TNTC TNTC ×
90 minutes × × × × ×
2) Pseudomonas aeruginosa 30 minutes TNTC TNTC TNTC TNTC TNT

C
60 minutes TNTC TNTC TNTC × ×
3) Bacillus subtilis 30 minutes TNTC TNTC TNTC TNTC TNTC
60 minutes × × × × TNTC
90 minutes × × × × ×
4) Escherchia coli 30 minutes TNTC TNTC TNTC TNTC TNTC
60 minutes TNTC TNTC TNTC TNTC TNTC
5) Candida albicans 30 minutes TNTC TNTC TNTC TNTC TNTC
6) Salmonella abony 30 minutes TNTC TNTC TNTC TNTC TNTC
90 minutes TNTC TNTC × × ×
7) E.I E.coli 30 minutes TNTC TNTC TNTC TNTC TNTC
34
8) E.I Bacillus subtilis 30 minutes TNTC TNTC TNTC TNTC TNTC
9) E.I Staphylococcus 30 minutes TNTC TNTC TNTC TNTC TNTC

aureus
*TNTC- Too numerous to count.
× = No growth.
OBSERVATIONS
Disinfectant Levermed.
Manufacturer Universal Healthcare Silvasa.
Manufacturing Date March 2008.
Expiry Date Feb 2010.
Batch no 803
Concentration Not available(ready to use)
35
Disinfectant Microbial culture Contact Dey-Engley broth tubes
time Tube 1 Tube 2 Tube 3 Tube 4 Tube 5
(minutes)
Levermed S.aureus 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
P.aeruginosa 3minutes √ √ √ √ ×
5 minutes √ √ √ √ ×
7 minutes √ √ √ √ ×
Bacillus sutilis 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
Escherchia coli 3 minutes √ √ √ √ √
5 minutes √ √ √ √ √
7 minutes √ √ √ √ ×
Candida 3 minutes √ √ √ √ √
5 minutes √ √ √ √ √
albicans
7 minutes √ √ √ × ×
Salmonella 3 minutes √ √ √ × ×
5 minutes √ √ √ × ×
abony
7 minutes √ √ √ × ×
E. I E. coli 3 minutes √ √ √ √ ×
5 minutes √ √ √ √ √
7 minutes √ √ √ √ √
E.I B. subtilis 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
E. I S. aureus 3 minutes √ √ √ √ √
5 minutes √ √ √ √ √
7 minutes √ √ √ √ √
Sign: √ = Growth. × = No growth.
Test Controls:
S.NO Control
Sign: √ = Growth.
× = No growth.
36
Enumeration of Inoculums on SCDA:
106 107 108 109 1010
2) Pseudomonas aeruginosa 50 9 2 × ×
4) Escherchia coli 240 16 2 × ×

6) Salmonella abony TNTC 18 3 × ×
7) Environmental isolates of Escherchia coli 102 25 2 × ×

8) Environmental isolates of Bacillus subtilis 100 25 2 × ×
9) Environmental isolates of Staphylococcus 103 26 3 × ×
aureus
Recovery:-
S.NO Microbial culture Contact time Cfu/mL(Dey –Engley tube)
(minutes) Tube 1 Tube 2 Tube 3 Tube 4 Tube
5
1) Staphylococcus aureus 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
2) Pseudomonas 3 minutes TNTC TNTC TNTC TNTC ×
aeruginosa 5 minutes TNTC TNTC TNTC TNTC ×
3) Bacillus subtilis 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
37
4) Escherchia coli 3 minutes TNTC TNTC TNTC TNTC TNTC
5) Candida albicans 3 minutes TNTC TNTC TNTC TNTC TNTC
6) Salmonella abony 3 minutes TNTC TNTC TNTC TNTC ×
5 minutes TNTC TNTC TNTC TNTC
7) E.I E.coli 3 minutes TNTC TNTC TNTC TNTC

×
8) E.I Bacillus subtilis 3 minutes × × × ×

×
5 minutes × × × ×
×
7 minutes × × × ×
×
9) E.I Staphylococcus 3 minutes TNTC TNTC TNTC TNTC TNTC
aureus
*TNTC- Too numerous to count, × = No growth.
38
OBSERVATIONS
Disinfectant OXIVIR.
Manufacturer Johnson Diversy.
Manufacturing Date August 2008.
Expiry Date August 2010.
Batch no 804
Concentration 2% v/v
39
Disinfectant time
(minutes) Tube 1 Tube 2 Tube 3 Tube 4 Tube
5
Oxivir S.aureus 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
P.aeruginosa 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
Bacillus subtilis 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
Escherchia coli 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
Candida albicans 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
Salmonella abony 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
E. I E. coli 3 minutes × × × × ×
5 minutes √ √ × × ×
7 minutes √ × × × ×
E.I B. subtilis 3 minutes √ √ × × ×
5 minutes √ √ × × ×
7 minutes √ × × × ×
E. I S. aureus 3 minutes √ √ √ × ×
5 minutes √ × × × ×
40
Sign: √ = Growth,× = No growth.
Test Controls:
S.NO Control
Sign: √ = Growth, × = No growth.
Enumeration of test inoculums on SCDA:

106 107 108 109 1010
3) Bacillus subtilis 115 67 3 × ×
5) Cabdida albicans 60 65 3 × ×

aureus
41
Recovery:
(minutes)
5 minutes × × × × ×
7 minutes × × × × ×
2) Pseudomonas 3 minutes × × × × ×
aeruginosa
5 minutes × × × × ×
7 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
4) Escherchia coli 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
5) Candida albicans 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
42
6) Salmonella abony 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
7) E.I E.coli 3 minutes TNTC TNTC × × ×
7 minutes TNTC × × × ×
8) E.I Bacillus subtilis 3 minutes TNTC TNTC × × ×
7 minutes × × × × ×
9) E.I Staphylococcus 3 minutes TNTC TNTC TNTC × ×

aureus
7 minutes × × × × ×
OBSERVATIONS
43
Disinfectant OXIVIR.
Manufacturing Date August 2008.
Expiry Date August 2010.
Batch no 804
Concentration 2.5% v/v

time
(minutes) Tube 1 Tube 2 Tube 3 Tube 4 Tube 5
Oxivir S.aureus 3 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
5 minutes × × × × ×
7 minutes × × × × ×
5 minutes × × × × ×
Test Controls:
S.NO Control
44

Enumeration:-
106 107 108 109 1010
3) Bacillus subtilis 115 67 3 × ×
5) Cabdida albicans 60 65 3 × ×

aureus
Recovery:-
45
(minutes)

60 minutes × × × × ×
90 minutes × × × × ×
aeruginosa
60 minutes × × × × ×
90 minutes × × × × ×

60 minutes × × × × ×
90 minutes × × × × ×
60 minutes × × × × ×
90 minutes × × × × ×

60 minutes × × × × ×
90 minutes × × × × ×

60 minutes × × × × ×
90 minutes × × × × ×
7) E.I E.coli 30 minutes × × × × ×

60 minutes × × × × ×
90 minutes × × × × ×
8) E.I Bacillus subtilis 30 minutes × × × × ×

60 minutes × × × × ×
90 minutes × × × × ×
46
9) E.I Staphylococcus 30 minutes TNTC × × × ×
aureus
60 minutes × × × × ×
90 minutes × × × × ×
47
OBSERVATIONS
Disinfectant VIREX II 256.

Manufacturing Date November 2008.
Expiry Date Novemner2010.
Batch no 804
48
time
Virex II 256 S.aureus 5 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
E. I E. coli 5 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
E.I B. subtilis 5 minutes √ × × × ×
10 minutes √ × × × ×
15 minutes × × × × ×
E. I S. aureus 5 minutes √ √ × × ×
10 minutes √ × × × ×
15 minutes
49 × × × × ×
Test Controls:
S.NO Control
Enumeration of test Inoculums on SCDA:

106 107 108 109 1010
1) Staphylococcus aureus TNTC 70 2 × ×

aureus
50
Recovery:-
(minutes)

10 minutes × × × × ×
15 minutes × × × × ×
aeruginosa
10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×
7) E.I E.coli 5 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×
8) E.I Bacillus subtilis 5 minutes TNTC × × × ×

15 minutes × × × × ×
51
9) E.I Staphylococcus 5 minutes TNTC TNTC × × ×
aureus
15 minutes × × × × ×
OBSERVATIONS
Disinfectant VIREX II 256.

Manufacturing Date November 2008.
Expiry Date Novemner2010.
Batch no 804
52
time
Virex II 256 S.aureus 5 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×
E. I E. coli 5 minutes TNTC × × × ×
15 minutes × × × × ×
E.I B. subtilis 5 minutes TNTC × × × ×
15 minutes × × × × ×
E. I S. aureus 5 minutes TNTC √ × × ×
15 minutes
53 × × × × ×
Test Controls:
S.NO Control
Enumeration of Test Strains on SCDA:

S. Microbial culture Cfu/mL
No
106 107 108 109 1010
1) Staphylococcus aureus TNTC 40 2 × ×
54
aureus
Recovery:-
(minutes)
Tube 1 Tube 2 Tube 3 Tube 4 Tube
5

10 minutes × × × × ×
15 minutes × × × × ×
aeruginosa
10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×
10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×

10 minutes × × × × ×
15 minutes × × × × ×
7) E.I E.coli 5 minutes TNTC × × × ×

15 minutes × × × × ×
55
8) E.I Bacillus subtilis 5 minutes TNTC × × × ×
15 minutes × × × × ×
9) E.I Staphylococcus 5 minutes TNTC × × × ×

aureus
10 minutes × × × × ×
15 minutes × × × × ×
Discussion
Disinfection in Pharmaceutical practice includes disinfection of the inanimate surfaces,
Instruments, articles used in aseptic processing. The process of disinfection removes most
of the harmful organisms but may not kill spores. Routine disinfection of surface has been
contaminated by infectious material or agents. Though cleaning of these surfaces may be
sufficient in non critical as up to 95% of harmful bacteria can be removed by good
cleaning with soap and water. However, it has been seen after cleaning the germs that have
been left behind soon begins to grow and accumulate. Therefore, use of disinfection in
critical and high risk areas like burn units and ICUs may be justified.
A sound cleaning and sanitization program is needed for controlled

environment used in the manufacture of pharmacopeial articles to prevent the microbial
contamination of these products. Sterile drug products may be contaminated via their
pharmaceutical ingredients, process water, packaging components, manufacturing
environment processing equipment and manufacturing operators. Current good
manufacturing practices (cGMP) emphasize the size, design, construction and location of
buildings and construction materials and the appropriate material flow to facilitate cleaning,
56
maintenance and proper operations for the manufacture of drug products. When disinfectants
are used in a manufacturing environment, care should be taken to prevent the drug product
from becoming contaminated form chemical disinfectant as a result of the inherent toxicity of
the disinfectants. The requirement for aseptic processing include readily cleanable floors,
walls and ceilings that have smooth and non porous surfaces; particulate, temperature and
humidity controls; cleaning and disinfecting procedures to produce and maintain aseptic
conditions. The cleaning and sanitization program should achieve cleanliness standards,
control microbial contamination of products, and be designed to prevent chemical
contamination of pharmaceutical ingredients; products contact surfaces or equipments,
packaging materials and ultimately the drug products. These principles also apply to non
sterile dosage forms where the microbial contamination is controlled by the selection of
appropriate pharmaceutical ingredients, utilities, manufacturing environments, sound
equipments cleaning procedures, products especially formulated to control water activity,
inclusion of suitable preservatives, and products packaging design.
In addition to disinfectants, antiseptics are used to decontaminate human skin and exposed
tissues and may be used by personnel prior to entering the manufacturing area.
Chemical sterilants mat be used to decontaminate surfaces in manufacturing and
sterility testing areas. Furthermore, sterilants may be used for the sterilization of
pharmacopeial articles, and UV irradiation may be used as a surface sanitizer.
The effectiveness of a disinfectant depends upon its intrinsic “biocidal” activity;
• The concentration of the disinfectant
• Contact time
• Nature of the surface disinfected
• Hardness of water used to dilute the disinfectant
• Amount of the organic material present on the surface
• Type and number of microorganism present
Under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA), the Environment
protection agency (EPA) registers chemical disinfectant marketed in the United States and
requires manufacturers to supply product information on the use dilution, and type of
microorganisms killed and the necessary contact time. Certain liquid chemicals sterilizers
intended for use on critical or semi critical medical device are defined and regulated by the
U.S Food and Drug Administration (FDA).Selection of an antiseptic for hand and surgical
site disinfection.
57
Disinfectants work by oxidizing the germs, breaking down their cell walls or by the otherwise
deactivating them. Different ingredients or combination of ingredients kill the different
germs, therefore, selection of the disinfectant will also depend upon the specific germs
required to kill. A broad spectrum disinfectant will be ideally suited.
A wide range of disinfectants is available commercially.
These includes phenols, Aldehydes, Quaternary ammonium compounds (QACs), chemical
based on release of nascent Chlorine, Iodophors, Hydrogen peroxide and combination of
various chemicals with detergents.
Though many products are commercially available, products and procedures described in
trade and other literature may not adequately decontaminate items when the surface has been
contaminated with highly resistant or unusual organisms, or if excessive numbers of
organisms are present. Disinfection is also affected by the quality of environmental hygiene
because dust and other organic matter present on the surfaces directly affect the efficacy of
disinfectant. When choosing a disinfectant for specific hospitals use it may be necessary to
know the number and the types of organisms likely to be present and the susceptibility of the
patients involved. Products and procedures described by manufacturer may therefore not
always the suitable for hospital use, and disinfectant chosen for one purpose may not be
equally effective if used for another.
Ideally the good disinfectant should have a broad antimicrobial spectrum and non–irritating,
non-toxic, non-corrosive and least expensive. Selection decision should include effectiveness
against potential pathogenic agent, safety to people, impact on equipment and the
environment and expense.
Although a few phenolic agents exhibit high toxicity and low biodegradability and as a result
has been banned, many of the phenolics are still in use throughout the world because of their
low cost. Phenol is rapidly corrosive on tissues. Keeping in view its less effectiveness and
potential health hazards, better and safe disinfectants are required to place them.
Chlorine has very efficient and rapid action on bacteria, viruses, and most fungi. But on the
other hand there are certain disadvantages. Those include:
 Its maximum effect is at pH-7, if pH is more than 8, the effect is strongly reduced.
 When pH falls down the stability of solution is reduced.
 The efficacy is strongly reduced by organic matter – when it reacts with proteins,
chloramines are produced which are bactericidal but work slowly.
 Chlorine preparations’ mixed with acid creates very toxic vapors.
 Strong solution of hypochlorites smell unpleasantly and are toxic–irritant to skin, eyes
and mucous memberane.
 Very corrosive, especially to aluminum but even stainless steel is also damaged. So if
used in modern pharmaceutical buildings for surface disinfection of floors and walls
these may corrode the aluminum doors and windows even if there are spashes of
chlorine solution on them.
58
Aldehyde containing disinfectants are widely used for the chemical sterilization of heat
sensitive equipment as they are non-corrosive to instruments, rubber and plastic articles. But,
Aldehydes cause irritation to skin, eyes. They may cause allergy, allergic asthma and allergic
contactexema. Inadequate rinsing of Gluteraldehyde causes Colitis. Keeping in view all the
above side effects, the health Authorities are considering the development and use of safer
alternatives to aldehydes. So it seems that the aldehyde based disinfectant is now discouraged
because of its harmful effects on humans, even when its antimicrobial activity is very good.
The Iodophors provide wide germicidal activity and show limited antimicrobial effects in the
presence of organic matter. They are corrosive and stain fabric and equipment. Traditionally
Iodine compounds have been used for skin disinfection purposes. Scanty literature is found
which shows the use of the “Tamed Iodine” compounds for surface disinfection including
instruments, walls and floors.
Among Quaternary ammonium compounds, two products are active against a wide range of
microorganisms including yeast and moulds. These are odorless; colorless; non-corrosive;
and highly stable compound over a wide range of pH (3-10.5) and temperature. They are
relatively stable in presence of organic matter, have bacteriostatic residual effect on treated
surfaces, caused low irritation and low toxicity.
In general disinfectants belonging to QACs group were discouraged because of their selective
varied antimicrobial efficacy, slow activity against Coliforms and gram negative bacteria, less
effectivity on spores and bacteriophages, and their effectivity is affected by hard water salts.
 Four locally available disinfectants were included for efficacy study: Levermed,
Incidur, Oxivir, Virex II 256. All the disinfectants were used at their working
dilutions as recommended by the manufacturer and as mentioned in manufacturer
literature.
Disinfectant efficacy and their shelf life were tested by In-Use Dilution test method. In this
method, we were screening disinfectants for their efficacy and shelf life at various
concentrations and contact time against a wide range of standard test organisms and
environmental Isolates.
For the In-Use Dilution method, the test organisms are enumerated on Soyabean-casein
Digest Agar by pour plate method. Neutralizers that inactivate the disinfectants should be
included in either the diluent or microbiological media used for the microbial enumeration.
Dey-Engley neutralizing broth is purposely used for disinfectant efficacy study.
In our present Study we found that, the average reduction of test microorganisms was found
to be Maximum by Virex II 256 and Oxivir. Both Virex II 256 and Oxivir found to be
59
effective against all standard microbial strain and environmental Isolates at manufacturer
recommended concentration and contact time.
Therefore, the results of present Disinfectant Efficay study shows that Gluteral, Glyoxal
(Incidur) and n-popano, Isopropyl, Benzalkonium chloride (Levermed) are not good non-
efficient disinfectants. On the other hand, Hydrogen peroxide, Alkyl Benzene sulphonic acid,
Alcohol ethoxylate, Hydorxyethene-1-1-diphosphonic acid, 2- butoxyethanol (Oxivir)and
Didecyl dimethyl ammonium chloride, benzyl ammonium chloride, amine oxide,, sodium
bicarbonate (Virex II 256) are good and efficient disinfectant in our findings. Phenolics and
Benzalkonium Chloride are not efficient disinfectants but their formulation with other
chemical derivatives increases their effectiveness. So, newer generation disinfectant such as
Oxivir and Virex II 256 are more effective than traditionally used phenolics and
Benzalkonium chlorides.
However, other Issues like cost, safety for humans use, and corrosiveness of metallic surfaces
need to be addressed before choosing an Ideal disinfectant for Pharmaceutical use.
BIBLIOGRAPHY
1. A Guide to selection and use of disinfectants, BC center for disease control, 2003.
2. Antiseptics and Disinfectants: Activity, Action, and Resistance, Clinical

microbiology reviews, 1999, p 147-179 Vol.12 No.1.
3. Advanced Sterilization Products, Johnson & Johnson. Oxivir OPA high level
disinfection solution: technical information. Irvine: Advanced Sterilization
Products; 1999.
4. AOAC International official methods of analysis, 15, 16 and 17 th editions,

Arlington, USA.
60
5. Ascenzi, J.M, Ed. Handbook of disinfectants and Antiseptics, 5th ed; Marcel
Delcker: New York 1995;
6. Ayliffe, G.A.J, Brightwell, K.M, Collins, B.S, and Cowbury, E.J.L (1969),
varieties of aseptic practices in hospitals and wards; page no 1117-1120.
7. Ayliffe GAJ, Collins BJ and Taylor LJ. Hospital acquired infection principles and
prevention 1990. Published by Butterworth Heinmenn Ltd. Guildford.
8. Block S.S and Lawrence A.C. In historical review. Disinfection, sterilization and
preservation 1968.
9. British Standard specification, number 541 (1939) with amendments dated 1943-
1961; Obtainable from British standard Institution, 28 Victoria street, London.
10. Block, S.S, Ed. Disinfection, Sterilization and Preservatives, 5 th ed; Lippincott
Williams and Wilkins publishers: Philedelphia, 2000.
11. CDC Guidelines for infection control in Dental Health-care Settings 2003,
Vol.52/RR-17.
12. Dee Groiffin Nots, 1988. Selection and use of disinfectants.
13. Dakin H.D 1915. The antiseptic action of Hypochlorites. Brit med Journal.
14. Denny, V.F Marsik, F.J Current practices in the use of Disinfectants within the
Pharmaceutical industry PDA Journal of pharmaceuticals Science and
Technology, 1997.
15. Fraise AP. Choosing disinfectants. J hospital Infect 1999.
16. Guidelines for Infection Control Practices, APIC Guidelines for selection and use
of Disinfectants. AJIC 1996; 24: 313-342.
17. Gershinfield L. 1957. Povidine iodine as a topical antiseptic. American Journal of

Surgery 94, 938-939.
18. Hugo WB, Rusell AD. Types of antimicrobial agents Principles and practices of
disinfection, preservation and sterilization, 3rd edition Oxford: Blackwell
scientific publications, 1999.
61
19. Infection Control Guidelines, Hand washing, Cleaning, Disinfection and
Sterilization in Health care. CCDR 24S8, December 1998: Health Canada.
20. Jacobs, WA. 1916. The Bactericidal properties of the quaternary salts of
Hexamethylenetetramine. Journal of Experimental Medicine 23, 563-568.
21. J Kennedy, J Bek. Selection and use of Disinfectants. Nebraska Cooperative

Extension 1998.
22. Journals of parentral science and Technology.
23. John J, Merians. Surface-active agents. Disinfection, sterilization and

preservation 2001.Editor Seymour S. block, 5th edition, Lippincott Williams and
Wilkins. A Wolters Kluwer Company, Philedelphia; 283-320.
24. Kelsey JC and Sykes G, 1969. A new technique for the assessment of
Disinfectants with particulars reference to their use in hospitals, Pharmaceuticals
Journal 202, 607-609.
25. Kelsey, J.C (1970). Disinfectants for hospitals use: an interim statement,
Brit.hosp.J.Service.
26. Kelsey, J.C and Maurer, I.M (1966), An In-use test for hospitals disinfectants.
27. Kronig B, Paul T.L. The chemical foundations of the study of disinfection and of
the action of poison. Zeitscrift jii Hygeine.
28. Public Health Laboratory Service Committee on the testing and evaluation of
Disinfectants in hospitals.
29. Parentrla Drug Association (PDA), Current Practice in the Validation of Aseptic
Processing Technical Report, No 24, J. Pharma. Science & Technology Report.
30. Russel, A.D; Hugo, W.B; Ayliffle, G.A.J, Ed. Principles and practices of
disinfection, Sterilization and Preservation and 3rd edition; Blackwell Science Inc:
London 1999.
62
31. Sykes G. 1958. Disinfection and Sterilization, Theory and practice 2nd ed. (1964),
London Champman and Mall.
32. Safety measures for general procedures. Manual for control of hospital associated
infections-standard operating procedure published by National AIDS Control
Organizations, Ministry of Health & Family Welfare, Governent of India, 1999.
33. Therapeutic Products Programme Guidelines, Minister of Health Canada, 1999

Edition.
34. US Food and Drug Administration (FDA) Center for Drug Evaluation and
Research (2003), Guidance for Industry Sterile Drug products produced by
aseptic Processing.
35. United States Pharmacopoeial Convention, Rockville, MD, <1116>

Microbiological Evaluation of Clean Rooms and other Controlled Environment,
in USP 27, NF 22 (2004).
36. WA Rutala, D J Weber. Infection Control: the role of disinfection and sterilization
.ICHE December 1999.
37. WA Rutala D J Weber. Draft Guidelines for Disinfection and Sterilization in

Healthcare Facilities. CDC Healthcare infection Control Practices Advisory
Committee 2001.
38. WHO Guidelines on hand Hygiene in Health care, 15 January 2006.
39. ASIAN EDITION 2008- USP 31/ NF-26, Volume 1, Page no 493-497, 693.
63

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REVIEW OF LITERATURE

An effective cleaning and disinfection program in aseptic processing areas of a Good

A good understanding of test methods used to assess disinfectant effectiveness is important.

JOSEPH LISTER & ANTISEPTIC SURGERY

Introduction: By the middle of the nineteenth century, post-operative sepsis infection

Joseph Lister (1827-1912)

GERM CONNECTION WITH WOUND SEPSIS

c) Skin disinfection test- which evaluate antiseptic properties on intact skin.

While the scientific application of disinfectants, sterilants, and preservatives is limited

• The application of disinfectant in the sterile pharmaceutical manufacturing

Microorganisms differ greatly in their resistance to disinfecting agents. The order of

1) Type of Microorganisms Examples

1) Bacterial spores Bacillus subtilis and Clostridium sporogenes.

2) Mycobacteria Mycobacterium tuberculosis.

3) Non lipid-coated viruses Poliovirus and Rhinovirus.

5) Vegetative bacteria Pseudomonas.aeruginosa,

6) Lipid-coated viruses Herpes simplex virus, Hepatitis B virus, HIV

Chemical Entity Classification Examples

2) Alcohol General purpose disinfectants, antiseptic, 70% Isopropyl

MECHANISM OF DISINFECTANT ACTIVITY

Cell wall Formaldehyde, hypochlorite’s and mercurials

MICROBIAL RESISTANCE TO DISINFECTANTS:

The development of microbial resistance to antibiotics is a well described phenomenon. The

2) Surface challenge test: - Using standard test micro-organisms and test

3) A statistical comparison of the frequency of isolation and numbers of microorganisms

TABLE: - NEUTRALIZING AGENTS FOR COMMON DISINFECTANT

Disinfectant Neutralizing Agents

1) Alcohols Dilution or polysorbate 80

Universal neutralizer’s broths may be formulated to contain a range of neutralizing agents.

TYPICAL CHALLENGE ORGANISMS

AOAC Challenge Organisms Isolates Typical Environmental

Bactericide: Escherchia. coli Bactericide: Micrococcus. luteus

Staphylococcus. aureus Streptococcus. Epidermis

Pseudomonas aeruginosa Coynebacteria.jeikeium

Fungicide: Penicillium. Chrysogenum Fungicide: C.albicans,

Sporicide: Bacillus. Subtilis Sporicide: Bacillus. Sphaericus

DISINFECTION IN CLEANING AND S A N I T I Z AT I O N

1) Nature of the item to be disinfected:

2) Number of microorganisms present:

4) Type and concentration of disinfectant used:

Resistance of microorganisms depends on the type of disinfectant used. A particular

The concentration of a particular disinfectant effects disinfection. In most cases, a higher

General Description of disinfectants used:

Incidur is a broad spectrum; Microbiocidal action including HIV, Hepatitis B etc.It

Active ingredients in 100 gms solutions:

 Commonly used for general purpose in laboratories and pharmaceuticals units.

 It is also responsible for general oxidation of cellular components in microbial cells.

Recommended concentration and contact time (According to manufacturer):

• According to manufacturer (Henkel Germany), Incidur is effective at 1%

Levermed is broad spectrum hand disinfectant. It acts as Bactericidal, Sporicidal,

Active Ingredients (w/w):

• Industrial Quality Control Laboratories, Research Institutes, Academics, Drug

• It generally shows action towards membrane permeability. It coagulates cellular

Recommended concentration and contact time (According to manufacturer):

• According to manufacturer (Universal Healthcare, Silvassa), Levermed is effective at

Oxivir is broad spectrum environmental cleaner & disinfectant. It is oxygen based

Alkyl Benzene sulphonic acid

Feature Advantage Benefits

Accelarated hydrogen Higher grade disinfectant Broad spectrum Microbial kill

Aldehyde free product No negative occupational Eco-friendly and user friendly

Cleaner and Single shot product Time and cost effective

 Oxivir is purposely used in Hospitals, laboratories, Institutes and Drug manufacturing

Recommended Concentration and contact time: