11.

3 Chemical Agents in
Microbial Control
Chemical control of microbes probably emerged as a serious
science in the early 1800s, when physicians used chloride
of lime and iodine solutions to treat wounds and to wash their
hands before surgery. At the present time, approximately
10,000 different antimicrobial chemical agents are manufac-
tured; probably 1,000 of them are used routinely in the health
care arena and the home. A genuine need exists to avoid in-
fection and spoilage, but the abundance of products available
to “kill germs,” “disinfect,” “antisepticize,” “clean and sani-
tize,” “deodorize,” “fight plaque,” and “purify the air” indi-
cates a preoccupation with eliminating microbes from the
environment that, at times, seems excessive (Insight 11.2).

Antimicrobial chemicals occur in the liquid, gaseous, or

even solid state, and they range from disinfectants and antisep-
tics to sterilants and preservatives (chemicals that inhibit the
deterioration of substances). For the sake of convenience (and
sometimes safety) many solid or gaseous antimicrobial chemi-
cals are dissolved in water, alcohol, or a mixture of the two to
produce a liquid solution. Solutions containing pure water as
the solvent are termed aqueous, whereas those dissolved in pure
alcohol or water-alcohol mixtures are termed tinctures.

Choosing a Microbicidal Chemical

The choice and appropriate use of antimicrobial chemical
agents is of constant concern in medicine and dentistry. Al-
though actual clinical practices of chemical decontamination
332 CHAPTER 11 Physical and Chemical Control of Microbes

TABLE 11.5 Qualities of Chemical Agents Used in Health Care

Agent Target Microbes Level of Activity Toxicity Comments
Chlorine Sporicidal (slowly) Intermediate Gas is highly toxic; Inactivated by organics;
solution irritates skin unstable in sunlight
Iodine Sporicidal (slowly) Intermediate Can irritate tissue; toxic Iodophors* are milder
if ingested forms
Phenolics Some bacteria, viruses, Low to intermediate Can be absorbed by skin; Poor solubility; expensive
fungi can cause CNS damage
Alcohols Most bacteria, viruses, Intermediate Toxic if ingested; a mild Flammable, fast-acting
fungi irritant; dries skin
Hydrogen peroxide,* Sporicidal High Toxic to eyes; toxic if Improved stability; works
stabilized ingested well in organic matter
Quaternary ammonium Some bactericidal, Low Irritating to mucous Weak solutions can
compounds virucidal, fungicidal membranes; poisonous if support microbial growth;
activity taken internally easily inactivated
Soaps Certain very sensitive Very low Nontoxic; few if any Used for removing soil,
species toxic effects oils, debris
Mercurials Weakly microbistatic Low Highly toxic if ingested, Easily inactivated
inhaled, absorbed
Silver nitrate Bactericidal Low Toxic, irritating Discolors skin
Glutaraldehyde* Sporicidal High Can irritate skin; toxic if Not inactivated by organic
absorbed matter; unstable
Formaldehyde Sporicidal Intermediate to high Very irritating; fumes Slow rate of action;
damaging, carcinogenic limited applications
Ethylene oxide gas* Sporicidal High Very dangerous to eyes, Explosive in pure state;
lungs; carcinogenic good penetration;
materials must be aerated
Dyes Weakly bactericidal, Low Low toxicity Stains materials, skin
fungicidal
Chlorhexidine* Most bacteria, some Low to intermediate Low toxicity Fast-acting, mild, has
viruses, fungi residual effects

*These chemicals approach the ideal by having many of the following characteristics: broad spectrum, low toxicity, fast action, penetrating abilities, residual
effects, stability, potency in organic matter, and solubility.

vary widely, some desirable qualities in a germicide have endospores, and, if properly used, are sterilants. Materials
been identified, including: that necessitate high-level control are medical devices—for
example, catheters, heart-lung equipment, and implants—
1. rapid action even in low concentrations,
that are not heat-sterilizable and are intended to enter body
2. solubility in water or alcohol and long-term stability,
tissues during medical procedures. Intermediate-level germi-
3. broad-spectrum microbicidal action without being toxic
cides kill fungal (but not bacterial) spores, resistant pathogens
to human and animal tissues,
such as the tubercle bacillus, and viruses. They are used to
4. penetration of inanimate surfaces to sustain a cumula-
disinfect items (respiratory equipment, thermometers) that
tive or persistent action,
come into intimate contact with the mucous membranes but
5. resistance to becoming inactivated by organic matter,
are noninvasive. Low levels of disinfection eliminate only
6. noncorrosive or nonstaining properties,
vegetative bacteria, vegetative fungal cells, and some viruses.
7. sanitizing and deodorizing properties, and
They are used to clean materials such as electrodes, straps,
8. affordability and ready availability.
and furniture that touch the skin surfaces but not the mucous
As yet, no chemical can completely fulfill all of those require- membranes.
ments, but glutaraldehyde and hydrogen peroxide approach
this ideal. At the same time, we should question the rather
overinflated claims made about certain commercial agents
such as mouthwashes and disinfectant air sprays.
Factors That Affect the Germicidal
Germicides are evaluated in terms of their effectiveness in Activity of Chemicals
destroying microbes in medical and dental settings. The three Factors that control the effect of a germicide include the na-
levels of chemical decontamination procedures are high, in- ture of the microorganisms being treated, the nature of the
termediate, and low (table 11.5). High-level germicides kill material being treated, the degree of contamination, the time
 11.3 Chemical Agents in Microbial Control 333

TABLE 11.6 Required Concentrations and volume of the liquid chemical (solute) is diluted in a larger
Times for Chemical Destruction volume of solvent to achieve a certain ratio. For example, a
of Selected Microbes common laboratory phenolic disinfectant such as Lysol is
usually diluted 1:200; that is, one part of chemical has been
Organism Concentration Time added to 200 parts of water by volume. Solutions such as
Agent: Aqueous Iodine chlorine that are effective in very diluted concentrations are
Staphylococcus aureus 2% 2 min expressed in parts per million (ppm). In percent solutions,
Escherichia coli 2% 1.5 min the solute is added to water by weight or volume to achieve
Enteric viruses 2% 10 min a certain percentage in the solution. Alcohol, for instance, is
Agent: Chlorine used in percentages ranging from 50% to 95%. In general, so-
Mycobacterium tuberculosis 50 ppm 50 sec lutions of low dilution or high percentage have more of the
Entamoeba cysts (protozoa) 0.1 ppm 150 min active chemical (are more concentrated) and tend to be more
Hepatitis A virus 3 ppm 30 min germicidal, but expense and potential toxicity can necessitate
Agent: Phenol using the minimum strength that is effective.
Staphylococcus aureus 1:85 dil 10 min Another factor that contributes to germicidal effective-
Escherichia coli 1:75 dil 10 min ness is the length of exposure. Most compounds require ade-
quate contact time to allow the chemical to penetrate and to
Agent: Ethyl Alcohol
act on the microbes present. The composition of the material
Staphylococcus aureus 70% 10 min
Escherichia coli 70% 2 min being treated must also be considered. Smooth, solid objects
Poliovirus 70% 10 min are more reliably disinfected than are those with pores or
pockets that can trap soil. An item contaminated with com-
Agent: Hydrogen Peroxide
mon biological matter such as serum, blood, saliva, pus, fecal
Staphylococcus aureus 3% 12.5 sec
material, or urine presents a problem in disinfection. Large
Neisseria gonorrhoeae 3% 0.3 sec
Herpes simplex virus 3% 12.8 sec amounts of organic material can hinder the penetration of a
disinfectant and, in some cases, can form bonds that reduce
Agent: Quaternary Ammonium its activity. Adequate cleaning of instruments and other
Compound
reusable materials ensures that the germicide or sterilant will
Staphylococcus aureus 450 ppm 10 min
better accomplish the job for which it was chosen.
Salmonella typhi 300 ppm 10 min
Agent: Silver Ions
Staphylococcus aureus 8 g/ml 48 h
Escherichia coli 2 mg/ml 48 h
(Continued from page 315)
Candida albicans (yeast) 14 mg/ml 48 h
he disinfection of the hospital likely failed because of the diffi-
Agent: Glutaraldehyde
Staphylococcus aureus
Mycobacterium tuberculosis
2%
2%
1 min
10 min
T culty with disinfecting rough surfaces, such as concrete block
walls, porous mats, and brushed concrete floors.
Herpes simplex virus 2% 10 min The problem with Salmonella was brought under control by
Agent: Ethylene Oxide Gas several measures. First, the concrete block walls of the stalls were
Streptococcus faecalis 500 mg/l 2–4 min painted with a special epoxy product, making them smooth and
Influenza virus 10,000 mg/l 25 h easy to clean. Second, the more porous mats were discarded and
Agent: Chlorhexidine replaced with smooth, solid rubber mats. The walls and mats were
Staphylococcus aureus 1:10 dil 15 sec then cleaned with detergent containing a quaternary ammonium
Escherichia coli 1:10 dil 30 sec product, and then effectively disinfected with sodium hypochlorite
(bleach). Surfaces must be completely cleaned of organic material
before the disinfection process is begun.
Hand scrubbing of the surfaces—including the walls, mats,
brushed concrete floors and drains—was found to be more effective
of exposure, and the strength and chemical action of the ger- than power washing. Hand cleaning and scrubbing likely increased
micide (table 11.6). Standardized procedures for testing the the amount of time that the Salmonella was exposed to the deter-
effectiveness of germicides are summarized in appendix C. gent and quaternary ammonium disinfectant.
The modes of action of most germicides are to attack the cel- Additional hand-washing stations were installed, and faculty,
lular targets discussed earlier: proteins, nucleic acids, the cell staff, and students were provided education on infection control
wall, and the cell membrane. measures.
A chemical’s strength or concentration is expressed in See: Tillotson, K. et al. 1997. Outbreak of Salmonella infantis infection in
various ways, depending upon convention and the method a large animal veterinary teaching hospital. J. Am. Vet. Med. Assoc.
of preparation. The content of many chemical agents can be (12):1554–1557.
expressed by more than one notation. In dilutions, a small
334 CHAPTER 11 Physical and Chemical Control of Microbes
Germicidal Categories According
to Chemical Group
Several general groups of chemical compounds are widely
used for antimicrobial purposes in medicine and commerce
(see table 11.5). Prominent agents include halogens, heavy
metals, alcohols, phenolic compounds, oxidizers, aldehydes,
detergents, and gases. These groups will be surveyed in the
following section from the standpoint of each agent’s specific
forms, modes of action, indications for use, and limitations.
The Halogen Antimicrobial Chemicals
The halogens are fluorine, bromine, chlorine, and iodine, a
group of nonmetallic elements, all of which are found in
group VII of the periodic table. Although they can exist in
either the ionic (halide) or nonionic state, most halogens exert
their antimicrobial effect primarily in the nonionic state, not
the halide state (chloride, iodide, for example). Because fluo-
rine and bromine are difficult and dangerous to handle, and
are no more effective than chlorine and iodine, only the latter
two are used routinely in germicidal preparations. These ele-
ments are highly effective components of disinfectants and
antiseptics because they are microbicidal and not just micro-
bistatic, and they are sporicidal with longer exposure. For
these reasons, halogens are the active ingredients in nearly
one-third of all antimicrobial chemicals currently marketed.
Chlorine and Its Compounds Chlorine has been used for
disinfection and antisepsis for approximately 200 years. The
major forms used in microbial control are liquid and gaseous
chlorine (Cl2), hypochlorites (OCl), and chloramines (NH2Cl).
In solution, these compounds combine with water and release
hypochlorous acid (HOCl), which oxidizes the sulfhydryl
(SOH) group on the amino acid cysteine and interferes with
disulfide (SOS) bridges on numerous enzymes. The resulting
denaturation of the enzymes is permanent and suspends
metabolic reactions. Chlorine kills not only bacteria and en-
dospores but also fungi and viruses. Chlorine compounds are
less effective and relatively unstable, if exposed to light, alka-
line pH, and excess organic matter.
Chlorine Compounds in Disinfection and Antisepsis Gaseous
and liquid chlorine are used almost exclusively for large-scale
disinfection of drinking water, sewage, and wastewater from
such sources as agriculture and industry. Chlorination to a con-
centration of 0.6 to 1.0 parts of chlorine per million parts of
water will usually ensure that water is safe to drink. This treat-
ment rids the water of most pathogenic vegetative microorgan-
isms without unduly affecting its taste (some persons may
debate this). In chapter 22, however, you will learn about path-
ogenic organisms that can survive water chlorination.
Hypochlorites are perhaps the most extensively used of all
chlorine compounds. The scope of applications is broad, in-
cluding sanitization and disinfection of food equipment in
dairies, restaurants, and canneries and treatment of swim-
ming pools, spas, drinking water, and even fresh foods.
Hypochlorites are used in the allied health areas to treat
wounds and to disinfect equipment, bedding, and instru-
ments. Common household bleach is a weak solution (5%) of
sodium hypochlorite that serves as an all-around disinfec-
tant, deodorizer, and stain remover.
Chloramines (dichloramine, halazone) are being em-
ployed more frequently as an alternative to pure chlorine in
treating water supplies. Because standard chlorination of
water is now believed to produce unsafe levels of cancer-
causing substances such as trihalomethanes, some water dis-
tricts have been directed by federal agencies to adopt
chloramine treatment of water supplies. Chloramines also
serve as sanitizers and disinfectants and for treating wounds
and skin surfaces.
Iodine and Its Compounds Iodine is a pungent black chem-
ical that forms brown-colored solutions when dissolved in
water or alcohol. The two primary iodine preparations are
free iodine in solution (I2) and iodophors. Iodine rapidly pene-
trates the cells of microorganisms, where it apparently dis-
turbs a variety of metabolic functions by interfering with the
hydrogen and disulfide bonding of proteins (a mode of ac-
tion similar to chlorine). All classes of microorganisms are
killed by iodine if proper concentrations and exposure times
are used. Iodine activity is not as adversely affected by or-
ganic matter and pH as chlorine is.
Applications of Iodine Solutions Aqueous iodine contains
2% iodine and 2.4% sodium iodide; it is used as a topical an-
tiseptic before surgery and occasionally as a treatment for
burned and infected skin. A stronger iodine solution (5% io-
dine and 10% potassium iodide) is used primarily as a disin-
fectant for plastic items, rubber instruments, cutting blades,
thermometers, and other inanimate items. Iodine tincture is a
2% solution of iodine and sodium iodide in 70% alcohol that
can be used in skin antisepsis. Because iodine can be ex-
tremely irritating to the skin and toxic when absorbed, strong
aqueous solutions and tinctures (5–7%) are no longer consid-
ered safe for routine antisepsis. Iodine tablets are available
for disinfecting water during emergencies or destroying
pathogens in impure water supplies.
Iodophors are complexes of iodine and a neutral poly-
mer such as a polyvinylalcohol. This formulation allows the
slow release of free iodine and increases its degree of pene-
tration. These compounds have largely replaced free iodine
solutions in medical antisepsis because they are less prone to
staining or irritating tissues. Common iodophor products
marketed as Betadine, Povidone (PVP), and Isodine contain
2% to 10% of available iodine. They are used to prepare skin
and mucous membranes for surgery and injections, in surgi-
cal handscrubs, to treat burns, and to disinfect equipment
and surfaces. Although pure iodine is toxic to the eye, a re-
cent study showed that Betadine solution is an effective
means of preventing eye infections in newborn infants, and it
may replace antibiotics and silver nitrate as the method of
choice.

Phenol and Its Derivatives
Phenol (carbolic acid) is an acrid, poisonous compound
derived from the distillation of coal tar. First adopted by
Joseph Lister in 1867 as a surgical germicide, phenol was the
major antimicrobial chemical until other phenolics with
fewer toxic and irritating effects were developed. Solutions
of phenol are now used only in certain limited cases, but
it remains one standard against which other phenolic disin-
fectants are rated. The phenol coefficient quantitatively com-
pares a chemical’s antimicrobic properties to those of phenol.
Substances chemically related to phenol are often referred to
as phenolics. Hundreds of these chemicals are now available.
Phenolics consist of one or more aromatic carbon rings
with added functional groups (figure 11.12). Among the
most important are alkylated phenols (cresols), chlorinated
phenols, and bisphenols. In high concentrations, they are cel-
lular poisons, rapidly disrupting cell walls and membranes
and precipitating proteins; in lower concentrations, they in-
activate certain critical enzyme systems. The phenolics are
strongly microbicidal and will destroy vegetative bacteria
(including the tuberculosis bacterium), fungi, and most vi-
ruses (not hepatitis B), but they are not reliably sporicidal.
Their continued activity in the presence of organic matter
and their detergent actions contribute to their usefulness.
Unfortunately, the toxicity of many of the phenolics makes
them too dangerous to use as antiseptics.
Applications of Phenolics
Phenol itself is still used for general disinfection of drains,
cesspools, and animal quarters, but it is seldom applied as a
medical germicide. The cresols are simple phenolic deriva-
tives that are combined with soap for intermediate or low lev-
els of disinfection in the hospital. Lysol and creolin, in a 1% to
3% emulsion, are common household versions of this type.
The bisphenols are also widely employed in commerce,
clinics, and the home. One type, orthophenyl phenol, is the
major ingredient in disinfectant aerosol sprays. This same
phenolic is also found in some proprietary compounds (Lysol)
often used in hospital and laboratory disinfection. One par-
ticular bisphenol, hexachlorophene, was once a common ad-
ditive of cleansing soaps (pHisoHex) used in the hospital and
home. When hexachlorophene was found to be absorbed
through the skin and a cause of neurological damage, it was
no longer available without a prescription. It is occasionally
used to control outbreaks of skin infections.
Perhaps the most widely used phenolic is triclosan, chem-
ically known as dichlorophenoxyphenol (see Insight 11.2). It
is the antibacterial compound added to dozens of products,
from soaps to kitty litter. It acts as both a disinfectant and an-
tiseptic and is broad-spectrum in its effects.
Chlorhexidine
The compound chlorhexidine (Hibiclens, Hibitane) is a com-
plex organic base containing chlorine and two phenolic rings.
Its mode of action targets both cell membranes (lowering sur-
11.3 Chemical Agents in Microbial Control 335
OH OH CH3 OH
CH3
Phenol o - cresol p- cresol
(basic aromatic
ring structure)
OH OH OH
Cl Cl
CH2
Cl Cl
Cl Cl Cl
Chlorophene Hexachlorophene
(a chlorinated phenol) (a bisphenol)
FIGURE 11.12 Some phenolics.
All contain a basic aromatic ring, but they differ in the types of
additional compounds such as Cl and CH3.
face tension until selective permeability is lost) and protein
structure (causing denaturation). At moderate to high concen-
trations, it is bactericidal for both gram-positive and gram-
negative bacteria but inactive against spores. Its effects on
viruses and fungi vary. It possesses distinct advantages over
many other antiseptics because of its mildness, low toxicity,
and rapid action, and it is not absorbed into deeper tissues to
any extent. Alcoholic or aqueous solutions of chlorhexidine
are now commonly used for handscrubbing, preparing skin
sites for surgical incisions and injections, and whole body
washing. Chlorhexidine solution also serves as an obstetric an-
tiseptic, a neonatal wash, a wound degermer, a mucous mem-
brane irrigant, and a preservative for eye solutions.
Alcohols as Antimicrobial Agents
Alcohols are colorless hydrocarbons with one or more OOH
functional groups. Of several alcohols available, only ethyl
and isopropyl are suitable for microbial control. Methyl alcohol
is not particularly microbicidal, and more complex alcohols
are either poorly soluble in water or too expensive for routine
use. Alcohols are employed alone in aqueous solutions or as
solvents for tinctures (iodine, for example).
Alcohol’s mechanism of action depends in part upon its
concentration. Concentrations of 50% and higher dissolve
membrane lipids, disrupt cell surface tension, and compro-
mise membrane integrity. Alcohol that has entered the proto-
plasm denatures proteins through coagulation, but only in
alcohol-water solutions of 50% to 95%. Alcohol is the excep-
tion to the rule that higher concentrations of an antimicrobial
chemical have greater microbicidal activity. Because water is
needed for proteins to coagulate, alcohol shows a greater mi-
crobicidal activity at 70% concentration (that is, 30% water)
336 CHAPTER 11 Physical and Chemical Control of Microbes

than at 100% (0% water). Absolute alcohol (100%) dehydrates

cells and inhibits their growth but is generally not a protein
coagulant.
Although useful in intermediate- to low-level germicidal
applications, alcohol does not destroy bacterial spores at
room temperature. Alcohol can, however, destroy resistant
vegetative forms, including tuberculosis bacteria and fungal
spores, provided the time of exposure is adequate. Alcohol is
generally more effective in inactivating enveloped viruses
than the more resistant nonenveloped viruses such as po-
liovirus and hepatitis A virus.

Applications of Alcohols Ethyl alcohol, also called ethanol

or grain alcohol, is known for being germicidal, nonirritat-
ing, and inexpensive. Solutions of 70% to 95% are routinely
used as skin degerming agents because the surfactant action
removes skin oil, soil, and some microbes sheltered in
deeper skin layers. One limitation to its effectiveness is the
rate at which it evaporates. Ethyl alcohol is occasionally
used to disinfect electrodes, face masks, and thermometers,
which are first cleaned and then soaked in alcohol for 15 to
20 minutes. Isopropyl alcohol, sold as rubbing alcohol, is
even more microbicidal and less expensive than ethanol, but
these benefits must be weighed against its toxicity. It must
be used with caution in disinfection or skin cleansing, be-
cause inhalation of its vapors can adversely affect the ner-
vous system.