How chemistry contributes to the

understanding of household and personal

?care products

Disinfectants
Antiseptic
Preservatives

Chemical disinfectants,
antiseptics, & preservatives
Are chemicals which have the ability to
destroy or inhibit the growth of
microorganisms, and are used for this
purpose

Other terms

Sepsis = the presence of a toxin or pathogenic m.o in

blood & tissue

Fungicidal & Fungistatic = chemicals used to kill or

stop the growth of the fungi

Bacteriostatic & Bactericidal = chemicals used to

kill or stop the growth of the bacteria

Virocidal = chemicals used to destroy the viruses

Sporacidal = chemicals used to destroy the spores

preservatives
Are chemicals included in pharmaceutical
preparations to prevent microbial spoilage
of the products

disinfection
is the process of removing pathogenic ===
m.o. from the surfaces of objects
reducing them to a level acceptable for a===
defined purpose
a level which is harmful neither to health===
nor to the quality of perishable products

Disinfectants levels of actions

High level disinfection===indicates
destruction of all m. o. but not necessarily
bacterial spores
complete killing == sterilization==
those which can kill spores are described
(as liquid sterilants =chemosterilant

Disinfectants levels of actions

Intermediate level disinfection
indicates destruction of all vegetative ===
. bacteria including T.B
but may exclude viruses & fungi =
and have little or no sporicidal activity=

Disinfectants levels of actions

Low level disinfection
destroy most vegetative bacteria fungi ===
& viruses
not include spores & resistant bacteria =

antisepsis
Destruction or inhibition of m.o. on living tissues ===
The effect is to limit or prevent the harmful result of =
infection
Applied to skin & mucous membranes=
they must be not toxic or irritating for skin=
mostly used to reduce the microbial population on skin=
prior to surgery or on hands to help prevent the spread
of infection by this route

Preservatives
Chemicals included in pharmaceutical===
preparations to prevent microbial spoilage of the
product

to minimize the risk of the consumer acquiring an =&

infection when the preparation is administered
it must be able to limit proliferation of m.o. that==
may be introduced unavoidably during manufacture
& the use of non sterile products (oral drops &
injections)

Preservatives
should kill any microbial contaminant =
introduced inadvertently during use in
sterile products (eye drops & multidose
injections(
must be not toxic in relation to the =
intended route of administration of the
product

Factors affecting the choice of antimicrobial

agent
properties of the chemical agent-1
microbial challenge-2
intended application-3
environmental factors-4
toxicity of the agent-5

The process of microbial killing or growth

properties
of the
chemical agent is basically
inhibition
using
antimicrobial
==== chemical reaction---- which influenced by

a- concentration
b- temperature
c- pH
d- formulation

Microbial challenge
the type of m.o. & the levels of microbial ==
contamination (bioburden( have a significant
effect on the outcome of chemical treatment

if the bioburden (concentration) is high***

long exposure time
high conc. required

---
---

m.o. vary in their sensitivity to the action of chemical

agents

vegetative bacteria---disinfectant***
should be capable of killing most
vegetative bac. Within reasonable contact
period
antiseptic & preservatives are also==
expected to have a broad spectrum
antimicrobial activity

Mycobacterium tuberculosis
equipments for respiratory investigations or ==
therapy can be contaminated & must be
disinfected to safe level

Bacterial spores
the majority of antimicrobial agents have no ==
useful sporicidal action with exception of
aldehydes***
hypochlorites***
some peroxygens***
Physical methods of sterilization

Fungi- === vegetative formsensitive

spores ---more resistant
===

Viruses ===enveloped
non enveloped
===

Relative resistance of microbes

Highest resistance
Bacterial endospores

Moderate resistance

Pseudomonas sp.
Mycobacterium tuberculosis
Staphylococcus aureus
Protozoan cysts

Least resistance

21

most vegetative cells

Fungal spores
enveloped viruses
Yeast
Protozoan trophozoites

Killing-Resistant Microorganisms
22

Endospore formers (endospores):

Bacillus spp. & Clostridium spp. are highly
heat- and chemical-treatment resistant.
Mycobacterium spp. are highly
chemical-treatment resistant.
Certain Pseudomonas spp. can actually
grow in certain disinfectants (i.e., in quats
and iodophores(.
Naked viruses (i.e., those lacking lipid
envelopes( often are resistant to chemical
disinfection.

Factors affecting the choice of antimicrobial

agent
properties of the chemical agent-1
microbial challenge-2
intended application-3
environmental factors-4
toxicity of the agent-5

intended application
intended application of an antimicrobial ==
agent whether preservative, antiseptic or
disinfectant will influence it's selection &
also affect it's performance
e.g. medical preparations ingredients in the
formulation may antagonize preservative activity

in disinfection of instruments the chemical==

must not have a harmful effect (corrosion, affect
clarity of lenses, or change texture(
Fabrics==adsorb QAC
Rubber==phenol
the disinfectant can only act if it is in contact**
with the item
access to all parts of an instrument is essential++
for small items total immersion in the++
disinfectant is required

Factors affecting the choice of antimicrobial

agent
properties of the chemical agent-1
microbial challenge-2
intended application-3
environmental factors-4
toxicity of the agent-5

environmental factors
organic matter can have a drastic effect on==
antimicrobial activity
adsorption
#
chemical inactivation
#
Reducing the conc. of active agent in solution--
Acting as a barrier for penetration of disinfectant-
Blood, body fluids, pus, milk (reduce the effectiveness
of antimicrobial agents even in small amounts)

toxicity of the agent

Some agents cause irritation to skin,==
eyes,, respiratory distress
Phenolics, formaldehyde, gluteraldehyde **
Covered containers
Rubber gloves
Face protection

Types of compounds

Types of compounds
acids & esters-1
alchohols-2
aldehydes-3
biguanides-4
halogens-5
heavy metals-6
H2O2& peracid compounds-7
phenols (carbolic acid(-8
surface-active-agents-9

acids & esters

Organic acids are considered
Benzoic acid
used alone or in combination**
used as the sodium salt (sodium benzoate(**
it's active substance is the non-ionized acid **
it is vital antifungal agent**

used in combination with other agent (salicylic **

acid( in superficial fungal infection
it is effective at relatively lower pH values**
preserve acidic food products**
conc. 0.5-0.1% are suitable for oral**
preparations

Sorbic acid
widely used preservative
Potassium salt(=potassium sorbate(
inhibit mold growth in acidic food **
(cheese(
its activity in decreasing pH**
pharmaceutical preservative for gums, **
mucilage, & syrups

Sulphur dioxid, sulphites, & metabisulphites

have extensive use as preservative in**
food
sodium sulphite & metabisulphite or**
bisulphite have a role acting as
preservative & antioxidants

Esters of p-hydroxybenzoic acid ==

(parabens(
are derivatives of benzoic acid =neutral pH
methyl, ethyl, propyl, & butyl esters**
a combination of esters is used for**
preparations which have two phases
methyl ester == water soluble
--
propyl,& butyl ester ==give
--
protection to the oil phase
pharmaceutical preservatives**
(emulsions, creams, & lotions(

parabens
inhibit mold growth in galenicala, liquid**
cosmetics, food, & shampoos
very effective against fungi**
less effective against bacteria**

alchohols

alchohols
Disinfectant & antiseptic
(ethanol & isopropanol(
.bactericidal = vegetative bact**
not sporicidal**
have cleaning action= degermination of skin**
.surface before injection by wiping out the m.o

Ethyl alcohol---water is essential for activity

bactericidal----60%-95%
disinfectant of skin, clean instrument &===70%
surfaces
virucidal---enveloped viruses===

combination with chlorhexidine or iodine =====

produce more active preparations
ethanol is popular in pharmaceutical==
preparations & cosmetics
propanol==bactericidal==
pre-operative skin treatment
preservative for cosmetics

**
**

Aromatic alkyl alcohols

preservatives (cosmetics & eye drops) ==

benzyl alcohol***
chlorbutol***
phenyl ethanol***
phenoxy ethanol***
bronopol***

aldehydes
Gluteraldehyde
Formaldehyde

antimicrobial==
sporicidal==

Gluteraldehyde
broad spectrum=
vegetative bac.= in minutes
spores = 3 hours
--

--

aqueous sol.= Cidex( w/v(2%==

cold liquid chemical sterilization of medical & surgical=
materials that cannot be sterilized by other methods

Formaldehyde = Formalin (37%(

disinfectant=
preserve biological specimens=
medical equipments=

gaseous state (vapor(=toxic &=

carcinogenic

aldehydes
highly toxic agents=
used as sterilants=
high-level disinfectants kill spores=
tissue preservatives=
inactivate viruses during vaccine preparation =

biguanides
Chlorhexidine
Alexidine
low-level disinfectant used in hospitals & dental=
offices & veterinary clinics
skin degerming agents in preoperative scrubs or=
oral rinse (skin & mucous membrane(
cannot kill spores or T.B. , limited antifungal=

Chlorhexidine
residual chemicals that remain active on=
surfaces for extended periods of time, providing
longer-lasting antimicrobial effects

peridex==

halogens
Chlorine
Iodine
Bromine
Fluorine

Chlorine

disinfectant in the form of gas==

Or
in combination with other chemical==
substances

wide spectrum (fungi & viruses(==

high-level (high concentration(---kill==
T.B.& spores
chlorine & chlorine compound cause ==
the production of hypochlorous acid
NaOCL+H2O ==HOCL+NaOH
strong oxidizing agent
high-level
---
antimicrobial activity is
depend on pH (5(

---
---

Chloramines==organic chlorine compounds

disinfectant & antiseptic
--release chlorine slowly
--water treatment
===

Iodine
skin antiseptic (surgical scrub( ==
.bac. Fungi, viruses, spores, T.B ==
insoluble in water==
combination of iodine & an organic --
molecule (solubilizing agent=surfactant(
==iodine released gradually
iodophores

Fluoride
antiseptic==
water treatment==
toothpaste==

Heavy metals
Copper, zinc, mercury, silver
germicides==
low-level disinfectant or antiseptic==
not sporicidal==
teeth fillings==
Thimerosol==vaccine preservative
silver nitrate eye drops
restricted==toxic

==
---

H2O2& peracid compounds

high-level disinfectant (25%(==
antiseptic (3%(==
cleansing of wounds & ulcers==
catalase in human cells

H2O2
H2O +
nescent oxygen
sporocidal==
contact lenses preservative==

Phenols (carbolic acid( & phenolics

&intermediate low-level disinfectant==
antiseptic

preservative==
Bactericidal, fungicidal, virucidal==
not sporicidal==
active at acidic pH==
caustic effect on skin==
toxic -- systemic==

the more highly substituted phenols are ==

less toxic & can be used as preservative &
antiseptic
-Carbolic acid
J. Lester( 1867(
Phenolics ==bactericidal, fungicidal, virucidal
not sporocidal
==
Lysol=cresol ==used widely household
antibacterial soaps
==
washing creams
==

surface-active-agents (surfactants)
anionic**
According to the ionization of the hydrophilic
cationic** group of the molecule
non-ionic**
Anionic= mainly used as sanitizers
Cationic=are quaternary ammonium compounds
( QUATS
(

effective at alkaline pH==

bactericidal (Gm.+ve bact(==

limited antifungal==
non sporicidal==
skin treatment, wound, abrasions & in ==
certain instances as preservatives

Benzyl cholium chloride

Cetrimide
surgery==
Aqueous & alcoholic solution & as creams
urology & gynecology==

Detergent
Dish soap
Household cleaners
Some mouth washes

Sites of action of chemical

antimicrobial agents

Sites of action of chemical antimicrobial agents

Chemical agents of control have 3 primary sites of action: cell wall & cellular
.membranes, proteins and nucleic acids (DNA or RNA)

Sites of action of chemical antimicrobial

agents
Disinfectants can act on m.o. in 2 different ways
growth inhibition (bacteriostatic , fungistatic( ##
or
lethal action (bactericidal,fungicidal, virucidal( ##

Disinfectants are usually complex formulation of

active molecules
Commonly used active ingredients in disinfectants and
antiseptics include
,organic phenolics
,quaternary detergents
, alcohols

. Iodophors= (iodine containing compounds(

Disinfectant and antiseptic compounds

that persist on a treated surface are
,termed residual, ie betadine

and
those compounds that evaporate or
degrade rapidly are non-residual, ie
. alcohols

:sometimes also containing

co-solvents--
chelating agents--
acidic or alkaline agent--
surface-active--
anti-corrosive products--

It should also be noted that may be

considerable variation in terms of
pH--
hardness--
salinity--

Possible stages of the mode of action

Primary stagecharacteristic of the mode

of action
Secondary stageonly a consequence of
action

Action on the external membrane of bacterial cell

membrane consist of basic compounds (phospholipids &-
lipopolysaccharides( , and is stabilized by Mg & Ca cations
if ionised disinfecting molecules are absorbed by electrical--
charges at the initial contact & absorption stage , the means of
action will be possible
non-polar molecules may dissolve & enter the lipid phase***

specific carrying system will lead other molecules through***

the membrane
other molecules will be able to disturb the organization of***
the membrane by remaining bound to certain sites

Action on the bacterial wall

bacterial cell wall is important & it differs--
, between Gm.+ve & Gm.-ve
which leads to great variation in the affinities of
the hydrophilic disinfectants

Action on the cytoplasmic membrane

an active molecule may penetrate the --

cytoplasmic membrane in 2 ways
Passive diffusion (non-specific & slow(**
Active transport (specific=enabling the **
accumulation of products in bacteria after

binding to membrane protein---

Action on energy metabolism

--some disinfectants acting on adenosine
triphosphatas (ATP( production

Action on the cytoplasm & nucleic acid

disinfectant mechanism may operate the --
cytoplasm & nucleic acid at the
chromosome level

Action on bacterial spores

spores are impermeable due to the --

presence of dipicolinic acid make them
more resistant
the presence of highly oxidizing product**
(chlorine( can destabilize this structure in
spores

action of various disinfectants

action of various disinfectants

Acidic & alkaline compounds
the efficacy of acidic & alkaline agents is linked to --
the concentration of

hydrogen (H(***
hydroxyl (OH( ions***
H ions # destroy the a.a. bond in nucleic acid
modify the cytoplasmic pH
#
precipitate proteins
#

OH ions # saponify the lipids in the enveloping

membrane
destruction of the superficial
structure
pH increase more than 10
disorganizes the structure of
peptidoglycan
causes hydrolysis of the
nucleotides of viral genome

#
#

pH exceed 12 # to act on Mycobacteria

Chlorine & derivatives

Chlorine is electronegative
oxidizes peptide links
*
denatures proteins
*
Hypochlorite & chloramine in water
hypochloric acid
Exposure to lethal dose of hypochloric acid
Decrease in ATP production
***
Permeability of external membrane

***

Exposure to sub-lethal dose

inhibit respiration due to a non-specific oxidizing effect ***

Quaternary ammonium compounds (QAC(

QAC irreversibly bind to the phospholipids &
proteins of the membrane

impairing permeability
the sensitivity is depend on the capacity of the bacterial--
cell to absorb such molecules
Gm.-ve.=lipoprotein & liposaccharides on the outer
membrane
Gm.+ve =wall protein
Thus able to enter & destroy membrane

in both Gm.-ve. & Gm.+ve. The--

antimicrobial activity of QAC with an alkyl
chain is related to lipophilia
QAC cause increase permeability

loss of viability

damage in outer membrane

metabolic injury & modification of
permeability

Phenolic compounds
Phenol acts specifically on the cell membrane &
inactivates intracytoplasmic enzymes by forming
unstable complexes
the lipophilic molecules are trapped by the--
membrane phospholipids
in low concentration**
nucleic acid is liberated out side
##
in high concentration**
denaturation of protein
##
lysis of the cell membrane
##

Peracetic acid & hydrogen peroxide

Peracetic acid oxidises & denaturates proteins & lipids of
m.o.
Disorganization of the membrane

Biguanides
Chlorhexidine =primary site of action is cytoplasmic
membrane
modification of membrane
permeability
this is due to electrostatic interaction of the##
chlorhexidine with the acid phospholipids in the
cytoplasmic membrane

Iodine compounds
The activity requires to know the behavior of
iodine in
aqueous
*
alcoholic solution
*
Iodine containing products == antiseptic#
iodine monochloride == disinfectant#
Iodine decrease the oxygen requirement of
aerobic m.o = interferes at the level of the respiratory
chain of the m.o. by blocking the transport of electrons
through electrophilic reactions with the enzymes of the
respiratory chain
interact with the proteins of the
=
.cytoplasmic membrane

Aldehyde compounds
Formoldehyde === acts on

protein by denaturation
nucleic acid by alkylation

##
##

the action is pH-dependent===working better at**

alkaline pH

Glutaraldehyde = same mode of action

Mode of action of virucidal disinfectants

The factors that to understand the action
presence of lipids in the virus
==
size of the virus
==

Group A= lipid-containing viruses (enveloped(

herpes, influenza, measles =susceptible
Group B= small non-lipid viruses (no envelope(

polio virus, HAV = resistance

Group C= large non-lipid (no envelope(

adeno virus

the presence of lipid is associated with ##

a high degree of susceptibility to all
disinfectants
the absence of lipid & small are##
associated with resistance to lipophilic
chemical agents
QAC, phenols, biguanides ==group A
Chlorine, iodine, oxidizing agents, some
aldehydes, strong acid & alkiline agents
== most viruses

86

87

Evaluation of non-antibiotic antimicrobial agents

microbial growth
Dynamics of disinfectants

Evaluation of non-antibiotic
antimicrobial agents
in the lab. & under favorable condition***
when a fresh medium is inoculated with a given##
number of bacterial cells
cell growth

Nutrients+ microbial cells

by the time***

cell growth + energy + reaction products

nutrients decrease & microbial growth increase

the population growth is monitored over a##

period of time
plotting the data will yield a typical bacterial ##
growth curve
it is difficult to graph population changes of ##
such enormous magnitude by using arithmetic
number
this why logarithmic scales are generally##
used to graph bacterial growth

Bacterial kinetic parameters

Lag phase==cells initially adjust to the new

medium until they can start dividing regularly by
binary fission

Exponential log phase

Bacterial growth becomes limited
stationary phase
decrease in nutrients & increases in metabolic west product

Until eventually they show loss of viability

death phase

Bacterial growth expressed as change in the

number of viable cells versus time
Generation time are calculated during the
exponential phase (generation time = is the time
required for a cell to double(
The cells are dividing regularly & are growing by
geometric progression
Cells divide at constant rate====

During the death phase the number of viable cells

decreases geometrically (exponentially(
Nutrients+ microbial cells = cell growth + energy + reaction products

Lag phase=no change

Growth phase
number of cells increase
=
nutrients decrease (due to utilization of nutrients(
=
Stationary phase=accumulation of west products, toxic
metabolite & inhibitory compounds = this effect the
condition (pH(
unfavorable environment

growth rat
death

forces that affect the microbial growth

??What are the forces that affect the microbial growth

Dynamic of disinfectants
Dynamics= study the forces acting upon a
body
Dynamics of microbial growth=a study of
environmental forces that act

either to promote

Or

.to impede the growth of m.o

The course of the destruction of population of

bacteria by chemical agent or other lethal agents,
can be followed by taking samples at intervals &
determining the number of survivors by making
colony count on a suitable nutrient medium
It is necessary to ensure that the action of the
chemical should be completely stopped when the
sample is taken either by the use of neutralizing
agent or by dilution
If this not done there will be continuing action &
mortality obtained will be grater than that which
existed at the moment of sampling

Bacteria usually die at constant rate

.E.g. 1 million bacteria treated for 1 min

has died
100000 left 90%
.if is treated for another min
died
10000 90%

time/min

Deaths/min

Number of
survivors

1000000

900000

100000

90000

10000

9000

1000

900

100

90

10

If the death curve

,is plotted logarithmically
the death rate is

constant
straight line

The factors that affect the disinfection process

many factors must be considered in the application of **
any physical or chemical agent to inhibit or destroy
. microbial populations
they are classified into two groups==

factors related to a killing agent (biocide(( 1(

factors related to organisms to be killed( 2(

Factors influencing antimicrobial action

Related to killing agent

Related to organisms to be
killed

the concentration of the- 1

chemical agent

the number of organism to- 1

be killed

the time during which the- 2

chemical acts

kind of organism- 2

the temperature at which- 3

the chemical acts

the physiological state of- 3

the organism
the environmental- 4
condition in which the killing
take place

The term concentration is applied to chemical

-agents
For example 0.1.% phenol will not kill E. coli, but
. will only prevent its growth
At 10 times this amount, (1% phenol (will kill in
. 20-30 minutes
. phenol will kill in 2-3 minutes 5%

 Sufficient time for contact for chemical reactions that

occur must be allowed.
This is described as the

dynamics of disinfection
1-The process is a gradual operation in which the
number of organisms killed in unit time is greater in the
beginning, but becomes less and less as the exposure
period is increased.
The log of number of the surviving organisms in unit .2
. volume plotted against time gives a straight line curve

time is inversely proportional to the concentration ##

, factor
i.e. the longer the time, the smaller the concentration
factor

Other factors
Temperature##
.pH##

Temperature --- rise in temp. increase the

biocidal activity of a disinfectant
Microbes killed more rapidly by the same ---
concentration of the disinfectant at elevated
.temp
At certain times a longer time at lesser temperature##
is preferred to prevent the deterioration of flavor, texture,
of the material

.pH
during the disinfection process a charge of ----
pH can effect
rate of growth ***

potency of the agent ***

ability of the disinfectant to combination ***

with sites on the bacterial cell surface

potency of the agent

if the agent is acid or base it's degree of ionization will==
depend on pH
in some compounds the active site is the non-ionized==
molecule while the ion is inactive

So- condition of pH which favor the formation of ions of

these compounds will also reduce this activity
in other cases activity of the compound is due to the==
ionized molecule

ability of the disinfectant to combination

with sites on the bacterial cell surface
before an antimicrobial agent can exert it's --
effect in a cell it must combine with that cell
pH affect the adsorption process--
as increase external pH
surface more negatively charged

the cell --

cationic biocide bind more strongly to the cell --

surface with consequent increase in activity

Standardization of disinfectants
evaluation of liquid disinfectant

Standardization of disinfectants
evaluation of liquid disinfectant
To standardize antimicrobial agents, several kinds of
properties are evaluated.

, the ability to prevent growth of organisms( 1(

, the ability to kill organisms already grown( 2(
the spectrum of activity or the types of organisms ( 3(
, affected by an agent
the toxicity, or the ability to be used in or on living ( 4(
medium

A number of different methods have been developed to

. measure the effectiveness of disinfectants
the most widely used method

phenol coefficient.===the killing power of a disinfectant

against a test organism, as compared to that of phenol under
. identical conditions
: The conditions which must be specified are as follows
organism used in the test--
, age of the culture--
, temperature of the test--
, time of action of disinfectant--
, presence and amount of organic matter in the culture--
, proportion of disinfectant to the culture--
, composition and reaction of the transfer medium--
temperature and time of the incubation of the transfer --
medium

A series of dilutions of phenol is prepared in tubes of sterile distilled **

. water

A similar series of dilutions of the disinfectant to be tested is **

. prepared
. The tubes are kept in a water bath at a temperature of 20C**

To each dilution tube0.5 ml of 24-hr broth culture of the test **

organism (selected strains ofStaphylococcus aureus,orSalmonella
typhosa) is added. At intervals of 5, 10 and 15 minutes, a standard
loopful I s transferred from each dilution tube to a corresponding tube
. containing 10 ml of sterile nutrient broth
, The subculture tubes are incubated at 37C for 24 to 48 hours**
.presence or absence of growth is noted **

The phenol coefficient is then calculated

as the ratio of the highest dilution of the disinfectant

##
killing the organism in 10 minutes (no growth in subculture
tube) but not in 5 minutes (growth in subculture tube) to the
corresponding dilutions of phenol that will kill under like
.conditions

The phenol coefficient of a disinfectant is of great

value in determining the dilution at which it can be
. used effectively

Preservatives

preservatives
Are chemicals included in pharmaceutical
preparations to prevent microbial spoilage
of the products

Micro-organisms can contaminate the***

pharmaceutical products during their
manufacture and this may lead to the spoilage
and rejection of the contaminated product or
may even cause impairment or death to the
consumer
Micro-organisms like bacteria and fungi are ***
the major problem of contamination

Pharmaceutical products consist of large variety of ingredients in a

. complex physicochemical state

. These are used in prevention, mitigation and diagnosis of diseases

Any pharmaceutical product has the potential to get contaminated with
bacteria, yeasts or moulds, which may affect their safety, efficacy or
.acceptability to the patient or consumer

Such products must meet

the requirements of the Good Manufacturing Practice
---
(GMP)

should be stable
--elegant during storage and use by addition of preservatives
and also by product design and by product formulation and
packaging

---

Sources of Contamination
Microbial contamination from
raw materials,
#Dry powders of natural origin #

, the manufacturing equipments#

, process operators#

packaging materials#

environment#

Water is the main source contamination #

Staff involved in manufacturing due to poor hygienic (skin,direct#

contact)
. wet areas such as drains and sinks#

dry surfaces like floors and walls contaminate the product #

Airborne contamination may be due to the product left open#
. to the air
. Containers and closures should be sterile#

Packaging materials like cardboard, plug, card liners, etc are#

greatly contaminated with bacterial and fungi spores

therefore it is necessary to maintain the quality at all the

stages of the process

Pharmaceutical product must meet the Good

Manufacturing Practices (GMP( for stable and safe
product
Sterilization should be maintained while handling the raw
materials and the final product at every stage of
. development, manufacture and use

Factors which affects the Microbial Spoilage of

Pharmaceutical Products
Micro-organisms grow at neutral pH, therefore acidic***
.or alkaline formulations are less susceptible to spoilage
Water is the most important cause of the survival and***
. growth of micro-organisms

Other factors includes osmotic pressure, nutrient and***

oxygen availability, surface tension, oxidation-reduction
balance, storage temperature and to a less extent
packaging design

combination of two or more preservatives are##

used to extend the range & spectrum of preservation
E.g.--- germall 115 has antibacterial activity but
combined with parabens shoes antibacterial &
antifungal activity
combination of antimicrobial preservatives may##
exhibit synergy == two compound exerts a greater
inhibitory effect == widen the antimicrobial spectrum
Eye drops & contact lens solutions

Factors affecting preservative activity

interaction with formulation components **
properties of the preservative (homogeneous , solubility( **

effect of containers**

glass---can expected to retain their

preservative contact if the closure is airtight
plastic---may penetrate through & interact with it
rubber---also react with many preservatives

corks---source of spores & molds

type of organism**

pH**

Sterility assurance

Sterile = no surviving organisms

Sterility testing== it is a test which assesses whether a
sterile pharmaceutical product is free from contaminating
m.o. by inoculation of the whole or part of that product
with a nutrient medium == direct inoculation

Random sampling is applied to products

In the final container to test whether it is adequately
protected against microbial spoilage (bacteria & fungus(
nasal & ophthalmic with aqueous bases --
injections----- multiple dose injections --
infusions--
implants--
syringes--
bandages--
dressings--
needles--
surgical instruments--

Sterility assurance==efficacy test==sterility test

membrane filtration*****

Oil, ointment, solid, soluble powder

filters
colony count
direct inoculation *****

solutions

WHAT IS THE ANTIMICROBIAL

?EFFECTIVENESS TEST
Antimicrobial Effectiveness Test
demonstrates the effectiveness of the
. preservative system in a product
A product Is inoculated with a controlled quantity
. of specific microorganisms
The test then compares the level of
microorganisms found on a control sample
versus the test over a period of 28 days

AET=antimicrobial effectiveness test

The AET is performed by spiking a panel of
challenge microorganisms (representing Grampositive cocci, Gram-negative bacilli, yeast, and
mold( individually into the product and
determining the log reduction of organisms at
prescribed time intervals to quantitatively
evaluate the effectiveness of the antimicrobial
preservative to prevent microbial proliferation
and/or kill the organisms

Parenteral
Chlorocresol (0.1%)Benzylalcohol (1.0%)
Cresol (0.3%)Phenylmercuric nitrate,
Phenol (0.5%)acetate or borate (0.002%)
Chlorbutol
Opthalmic
Chlorhexidine acetate or gluconate (0.1%)Thiomersal (0.1%)
Benzalkonium chloride (0.3%)Chlorbutol (0.5%)
Oral liquids
Methyl, ethyl and propyl p-hydroxybenzoate
(parabenz) (0.3%)
Benzoic and sorbic acid (0.3-0.5%)
Chloroform (0.25%)
Bronopol (0.5%)
Creams
Parabenz (0.3%)Cetyltrimethylammonium
Chlorocresol (0.1%) bromide (1.0%)
Bronopol (0.5%)Phenylmercuric nitrate
(0.01%)