Chapter II

Review of Related Literature

This chapter contains literatures in line with the study. This chapter also presents additional
background information and data about variables used to support the study.

Akapulko

Akapulko (Cassia alata) is a legume that grows in some parts of Asia. The tendrils are
forked. The leaves are simple and spiral. The seeds are also used as cooking oil which can
externally calm itchiness and as wine which can cure stomach aches. Akapulko have the ability
to elaborate a ribosome-inhibiting protein called tricosanthin, which has displayed encouraging
signs as an anti-Human Deficiency Virus (HIV) drug. (Wiart, 2006).

Antibacterial

Antibiotic is anything that destroys bacteria or suppresses their growth or their ability to
reproduce. Heat, chemicals such as chlorine, and antibiotic drugs all have antibacterial properties.
Many antibacterial products for cleaning and hand-washing are sold today. Such products do not
reduce the risk for symptoms of viral infectious diseases in otherwise healthy persons. This does
not preclude the potential contribution of antibacterial products to reducing symptoms of bacterial
diseases in the home. (www.medicinenet.com/script/main/art.asp?articlekey=10215).

Staphylococcus epidermidis
According to Ogston (1884), S. epidermidis (Staphylococcus epidermidis) remains one of
the most significant pathogens causing disease in animals and human and methicillin-resistant S.
aureus (MRSA) is ranked among the most important and common pathogens resistant to multiple
antibiotics all over the world. The term S. aureus means Golden Cluster Seed and it is also called
as golden staph. This microorganism was first isolated in 1884 by Scottish surgeon (Sir
Alexander Ogston) from surgical abscesses (Ogston, 1884).. Rosenbach, a year later, used the term
taxonomically and therefore provided the first-ever account on genus Staphylococcus. The clinical
importance of this microorganism in humans and animals has been known since its isolation.

Bioassay
A bioassay is routinely known as a biological assay. Bioassays are mainly used to measure
effects of a matter on a living organism and are absolutely vital in the development of new drugs
and in supervising ecological pollutants. It is a procedure for the conclusion of the engrossment of
a particular constitution of a blend. Bioassay procedures can work out the engrossment of purity
or biological activity of a matter such as vitamin, hormone, and plant development component.
Bioassays may be qualitative or quantitative. Qualitative bioassays are used for assessing the
physical effects of a substance that may not be quantified, such as abnormal development or
deformity. Quantitative bioassays mean the estimation of the concentration or potency of a matter
by estimation of the biological answer that it makes. Quantitative bioassays are normally analyzed
using the methods of biostatistics. (www.omicsonline.org/bioassay-peer-reviewed-open-access-
journals.php)

Antioxidant

Antioxidant is a substance that prevents or slows the breakdown of another substance by

oxygen. Synthetic and natural antioxidants are used to slow the deterioration of gasoline and
rubber, and such antioxidants as vitamin C (ascorbic acid), butylated hydroxytoluene (BHT), and
butylated hydroxyanisole (BHA) are added to foods (seefood additives) to prevent them from
becoming rancid or from discoloring. (The Columbia Encyclopedia, 2015)

Phytochemicals
Divya et al. (2014) point out that phytochemicals are complex chemicals found in plants,
like in fruits and vegetables. They are not classified as vitamins, minerals or macronutrients, but
they contribute greatly to health and wellbeing. There are over 10,000 of them and they have
effects such as antioxidants, boosting the immune system, anti-inflammatory, anti-viral,
antibacterial and cellular repair. Phytochemicals can also prevent oxidation; they are
phytochemicals with antioxidant properties and commonly came from fruits or vegetables with
bright color.
Phytochemicals are chemical compounds that occur naturally to plants. Some are
responsible for color and other organoleptic properties of plants and may have a biological
significance, for example, carotenoids or flavonoids, but are not established as essential nutrients.
(https://en.wikipedia.org/wiki/Phytochemical)
In order to identify the phytochemical compounds fount in a plant material, phytochemical
screening is being conducted. Phytochemical screening includes extraction of the plant material.
Fresh plant tissues that will be used for phytochemical analysis should be dried before extraction.
It is also necessary that the drying should be under controlled conditions, to avoid too many
chemical changes on the material. Then extraction takes place using various methods like the use
of rotary evaporator or Soxhlet apparatus. (Raaman, N. 2006)
In investigating the complete phytochemical profile of a given plant species, fractionation
of a crude extract is needed in order for the constituents to separate prior to the chromatographic
analysis. The procedures that can be applied in determining a plant materials chemical
constituents are, procedure based on varying polarity, Thin layer Chromatography (TLC), Paper
Chromatography (PC), Gas liquid Chromatography (GLC), and High Performance Liquid
Chromatography (HPLC). (Harborne, J. B. 1198)

Saponins
Saponins are oily glycosides with foaming characteristics found in plant skin where they
form a waxy protective coating. Chemically, saponins consist of a polycyclic aglycone part,
sapogenin, is either steroid or a triterpenes. The foaming ability of saponins is called by the
combination of hydrophobic (fat-soluble) sapogenin and a hydrophilic (water-soluble) sugar part.
Saponins have bitter taste and not poisonous to warm-blooded animals. However, they can dissolve
red blood cells when injected directly into the bloodstream. They are mostly found in most
vegetables, beans and herbs (Microsoft Encarta, 2001)
 Saponins have medical values. They seem to reduce blood cholesterol levels by binding
with bile salt and cholesterol in the intestinal tract to facilitate its absorption. Studies have shown
that saponins also prevent colon cancer. By reacting with the cholesterol-rich membranes of cancer
cells, saponins limit growth of cancer cells. Plant produces saponins to fight parasitic infections.
Once ingested by human, saponins also seem to help immune system and to protect against viruses
and bacteria.
Saponins have detergent properties and are used commercially as cleaning agents and as
flame producers in fire-extinguishing fluids. Some of the sugars and saponins are useful as raw
materials for the synthesis of steroid hormones (Encyclopedia Britannica, 2006).

Tannins
Tannins, also called tannic acid, are amorphous and crystalline vegetable products obtained
from various plants. Tannins are any large polyphenolic compound containing sufficient hydroxyl
and other carboxyl groups to form strong complexes with proteins and other macromolecules.
They have molecular weights ranging from 500 to over 20,000. Tannins have variable composition
and are usually divided into hydrolysable tannins and condensed tannins (proanthocyanidins).
Hydrolysable tannins are esters of glucose or some other sugar with one or more trihydroxy-
benzoic acids. They are hydrolyzed by weak acids or weak bases to produce carbohydrate and
phenolic acids. Condensed tannins are also called proanthocyanidins. They are phenols of
moderately complex structure. They consist of polymers of 2 to 50 (or more) flavonoids units that
are joined by carbon-carbon bonds, which cannot be cleared by ydrolysis. Some very large
condensed tannin is insoluble while hydrolysable tannins and most condensed tannins are water-
soluble (Microsoft Encarta Premium, 2007).
Tannins have yellow-white to brown colour deepens when exposed to light. They taste
bitter and are astringent with a faint characteristic odour. Water, acetone, and alcohol readily
dissolve tannins, but benzene, ether and chloroform do not. Tannins bind and precipitate protein,
gelatin, and alkaloidal metallic salts.
The precipitate properties of tannins leads to their use of tanning agents for ik, dyeing
clothes, coagulants for rubber and in clarifying wines and beer. Tannic acid is invaluable as an
external medicine because it is astringent and antiseptic (Sackheim and Lehman, 2002).

Alkaloids
Alkaloids attempt assorted and important physiological effects on humans and other
animals. Generous alkaloids include morphine, strychnine, quinine, ephedrine, and nicotine. They
are cheap exposed to in plants and are especially common in certain families of flowering plants.
It has been suggested that they are simply waste products of plants metabolic processes, but
evidence suggests that they may serve specific biological functions. In colorful plants, the attention
of alkaloids increases by oneself former to announce formation and then drops off when the seed
is ripe, suggesting that alkaloids may play a role in this process. Alkaloids may except for protect
some plants from destruction by certain insect species. (www.britannica.com/science/alkaloid)
Flavonoids
Flavonoids are a large family of polyphenolic compounds synthesized by plants, they are
the most significant plant pigments for flower coloration, producing yellow or red/blue
pigmentation in petals designed to captivate pollinator animals. In higher plants, they also act as
chemical messengers, physiological regulators, and cell cycle inhibitors.
(lpi.oregonstate.edu/mic/dietary-factors/phytochemicals/flavonoids)

Glycosides
Glycosides are one of the wide varieties of naturally occurring substances. Most of them
are often found in plants, often as flower and fruit pigments. Various medicines, condiments, and
dyes from plants occur as glycosides. Several antibiotics are glycosides (e.g., streptomycin).
Saponins, widely distributed in plants, are glycosides that lower the surface tension of water;
saponin solutions have been used as cleansing agents. (www.britannica.com/science/glycoside)

Thin Layer Chromatography

Thin Layer Chromatography (TLC) is a technique used to monitor the reactions of a
substance toward another substance, and to determine the proper solvent system for performing
separations using column chromatography. TLC usually uses alumina or silica, which is in
stationary phase that are polar (standard) or non-polar (reverse phase). A mobile phase may also
be used in TLC, they are solvents whose polarity is being chosen.
In most laboratory applications, you will use the standard phase silica plates. The reaction
mixture will be applied to the solution in the plate then let the plate with solvent (combination of
solvents) to run and move through capillary action. The compounds will travel in different
distances up the plate, depending on their polarity. More polar compounds will "stick" to the polar
silica gel and travel short distances on the plate, while non-polar substances will diffuse into the
solvent and travel large distances on the plate. The measure of the distance a compound travels is
called Rf. This number, between zero and one, is determined by measuring the distance the
compound moved from the baseline (where it was originally spotted) divided by the distance the
solvent moved from the baseline. (Franz, K. J. et al. 2013)

Antibacterial Ointment
Mupirocin (Bactroban) ointment and retapamulin (Altabax) have not been studied in head
to head trials in the treatment of impetigo, so it is unclear if retapamulin (Altabax) is more effective
than mupirocin. Retapamulin (Altabax) is not FDA-approved for use in infections caused by
MRSA. At this time, retapamulin has only been compared to placebo. Retapamulin (Altabax) has
an advantage in that its dosage regimen is twice daily versus that of mupirocin, which is three
times daily; however, total treatment area for retapamulin should not exceed 100 cm2 in adults or
two percent of total body surface area (BSA) in children and adolescents. Retapamulin (Altabax)
is an alternative to mupirocin ointment for the topical treatment of impetigo due to S.aureus
(methicillin-susceptible isolates only) and S. pyogenes. Impetigo is usually a self-limiting skin
infection, but resistance patterns should be taken into account in the choice of therapy. ,
Mupirocin (Bactroban) cream is FDA approved for the treatment of secondary infected
traumatic skin lesions due to susceptible strains of S. aureus and S. pyogenes. It is not indicated
for impetigo; however, mupirocin ointment is FDA approved for the treatment of impetigo due to
S.aureus and S. pyogenes. Mupirocin (Bactroban) cream is not in a polyethylene glycol (PEG)
base like mupirocin (Bactroban) ointment. PEG can be absorbed from open wounds and damaged
skin therefore should be avoided in patients with moderate to severe renal impairment. Direct
comparative trials of the cream and ointment formulations are lacking, and they are not considered
interchangeable.
pH Level
Hydrophilic ointment formulations containing salicylic acid were prepared at different pH
levels by inclusion of phosphate buffers. Percutaneous salicylic acid absorption was studied by
measuring salicylate blood levels in rabbits at 1.5-hr. intervals from 1.5 to 7.5 hr. after ointment
application. The effect of 15% dimethyl sulfoxide in the various formulations was also determined.
Formulations without dimethyl sulfoxide produced maximal blood concentrations at the highest
pH level (10.78) and higher concentrations at the lowest pH (2.97) than at the intermediate ones
(4.48, 6.80, 9.23). Proportionally higher concentration of undissociated species appeared to
account for the increased absorption observed at the most acidic pH, while increased dissolution
was probably responsible for the greater blood levels recorded at the most alkaline pH. Dimethyl
sulfoxide produced a more rapid rate of salicylate absorption as well as greater peak blood levels
at each pH. However, its influence on rate of absorption and on peak blood levels was less
pronounced at the highest pH levels. This finding seems to indicate that the positive effect of
dimethyl sulfoxide on percutaneous salicylate absorption is due in part to its ability to enhance the
dissolution of salicylic acid.