COMPARISON OF EFFICACY AND IMPROVEMENT OF HYPOCHLOROUS ACID

BASED ANTISEPTIC AND CHLOROXYLENOL BASED ANTISEPTIC AGAINST

GRAM NEGATIVE BACTERIA

BACKGROUND KNOWLEDGE

Antiseptics are chemical compounds that are applied on the skin and tissue (open wounds) to kill
microorganisms such as bacteria, viruses and fungi(Joseph Lister’s Antisepsis System | Science
Museum, n.d.). They are used extensively in a variety of settings. Antiseptics find application in
hospitals, health care settings and at home for domestic purposes. Antiseptics was first invented
by Joseph Lister in 1864, he came across Louis Pasteur germ theory at Glasgow University while
working as Professor of Surgery. In Louis theory he had suggested that disease causing
microorganisms could be destroyed by exposing the infected area to germ killing chemicals.
Joseph Lister decided to apply this knowledge in the treatment of infections that occur after
surgery. He researched on the various methods he could prevent microorganisms from entering a
wound by creating a chemical barrier. He named the chemical substance an antiseptic. He used
carboxylic acid which killed germs on contact. In the 21st century different types of antiseptics
have been developed. They are known to contain different active ingredients. We have phenol
based antiseptics, alcohol based, quaternary ammonium based, diguanides based, peroxide based,
quinoline based and iodine based antiseptics. The different base ingredients give the antiseptic
different properties and different efficacy levels against different types of bacteria. The wide
spread use antiseptic has led to some bacteria mutation therefore causing the bacteria to be
resistant against certain antiseptics. Different antiseptics have different mechanism of action
against bacteria. Mechanisms involved include cross linking of proteins, membrane damage
involving phospholipid bilayers, leakage, uncoupling and DNA strand breakage(McDonnell &
Russell, 1999). The mechanism action depends on different factors such as concentration of the
antiseptic agent being used, temperature under which the bacteria is exposed to, kind and the
number of microorganisms being investigated. The efficacy of antiseptics vary with the
concentration and ingredients used during the production period. Some ingredients used tend to
be more effective against certain kind of bacteria than others. When the concentration is varied it
can either increase or decrease the efficacy of the antiseptic.
ABSTRACT

STATEMENT OF A PROBLEM

Antiseptics are chemicals used for cleaning wounds and skin. They either kill or prevent the
growth of bacteria. They do this through the disruption of the cell wall of microorganisms or
disrupting their metabolism. The wide spread use of antiseptics against bacteria has rendered
most of these bacteria resistant to different types of antiseptics, especially gram negative
bacteria. Examples of these bacteria include (Pseudomonas, Escherichia and Salmonella). The
named above bacteria have the ability to infections that are detrimental to the human health.
Such infections include meningitis, pneumonia and bloodstream cause infections. The cause for
this resistance to antiseptics include; an impermeable outer membrane which makes it difficult to
destroy. Horizontal gene transfer reduces the susceptibility of the bacterium to the effect of the
antiseptic. Bacteria are constantly subjected to different environmental conditions such as
extreme Ph, temperature, osmotic pressure and oxidizing agents. Oxidizing agents are known to
cause damage to cells of microorganisms. Among the most widespread oxidizing agents, reactive
oxygen species (ROS), such as hydrogen peroxide, and reactive chlorine species (RCS), such as
hypochlorous acid, are the most abundant.(Da Cruz Nizer et al., 2020). Sodium hypochlorite is
an oxidizing agent and is commonly used as a surface disinfectant rather than an antiseptic. As a
surface disinfectant it is a first acting antimicrobial agent. It has the ability to interact with
several biomolecules and destroy them. Hypochlorous acid has a small shelf life due to its
decomposition into oxygen and chlorate. Phenol based antiseptics such as chloroxylenol are most
effective against gram positive bacteria but less effective against gram negative bacteria and
viruses. It is mostly used in wound cleansing applications Chloroxylenol does not cause skin
irritation or pose the danger of corroding surfaces onto which it is applied unlike hypochlorous
acid which corrodes surfaces and cause skin irritation. Chloroxylenol is also more stable and has
a longer shelf life compared to hypochlorous acid. In this research project we review the
production of hypochlorous and chloroxylenol antiseptic. Comparison on the efficacy of both
antiseptics on gram negative bacteria will be studied and finally will try to improve the efficacy
of chloroxylenol against gram negative bacteria by reacting it with hypochlorous acid.
OBJECTIVES

1. To produce chloroxylenol based antiseptic and hypochlorous acid.

2. To measure and compare the efficacy of chloroxylenol and hypochlorous acid on gram
negative bacteria.
3. To evaluate the efficacy of (HOCl + chloroxylenol) on gram negative bacteria.

HYPOTHESIS

 There is no difference between the production of chloroxylenol based antiseptic and

hypochlorous acid.
 There is no difference in the efficacy levels of chloroxylenol and hypochlorous acid on
gram negative bacteria.
 There is no difference in efficacy between (HOCl and chloroxylenol), HOCl and
chloroxylenol.

JUSTIFCATION

The rise of gram negative resistance bacteria to various antiseptic has prompted for action to be
taken in-order to solve the issues pertaining to this. Gram negative bacteria are known to cause
various infections such as meningitis, blood related infections and pneumonia. Research in this
area is still at its young stage. Studies from(da Cruz Nizer et al., 2020) has shown that gram
negative bacteria have an impermeable membrane that is not easily dissolved under normal
acidic or conditions. Hypochlorous acid has the ability to penetrate the cell wall of the bacteria
through passive diffusion which results in negative effects on cellular macromolecules and
membrane related processes due to its strong oxidizing ability.

Hypochlorous acid is non-toxic to biological tissues and it is also safe to use around the eyes,
mouths, ears and other skin surfaces. However HOCl has the tendency of causing a drying effect
and irritation after application on the skin and wounds. In addition it has a first decomposition
rate, HOCl cannot be stored or used for long periods of time. It dissociates into H+ and OCl- ions
which dramatically reduces its concentration thus possibly lowering the overall effectiveness of
the compound.

Chloroxylenol is mostly effective against gram positive bacteria. Limited studies have been done
on its effectivity against gram negative bacteria. Chloroxylenol is skin friendly and has a longer
shelf life when compared to hypochlorous acid.

Because of this there is a need to develop alternative ways of dealing with the resistant gram
negative bacteria. This research project will first test the efficacy of the two base ingredients
(HOCl and chloroxylenol) on gram negative bacteria and then determine whether an
improvement can be made on either compounds to improve their properties or efficacy levels.