Antibacterial Activity of Two Herbs Phytochemical's

Pharmacology and Alternative Medicine Academic Journal
(PAMAJ); Vol.5, No.3, 2020;

ISSN (7392 – 5820);
p –ISSN 7880 - 7492
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ANTIBACTERIAL ACTIVITY OF TWO HERBS PHYTOCHEMICAL’S

USED IN MANAGEMENT OF SKIN DISEASES.
1
Ogbuanu, C.C., 1Amujiogu, S.N. and 2Ogunbodede, T.T.
1
Department of Industrial Chemistry, Enugu State University of Science and Technology, Enugu Nigeria. P.M.B.
01660, Enugu, Nigeria
2
Department of Applied Microbiology and Brewing, Enugu State University of Science and Technology, Enugu
Nigeria. P.M.B. 01660, Enugu, Nigeria
Abstract: Skin diseases have been of major concern recently due to their aching, scares and pain association with it. Due
to the hot climate and overcrowding households common in Nigeria, the populace are more prone to a wide range of fungal
skin conditions. This study was carried out to access and document the antibacterial activity of phytochemicals present in
two medicinal plants used in Oghe traditional medicine to management skin diseases. Simple chemical tests were
undertaken to test for the phytochemicals while disk diffusion method was employed for the antibacterial activities of the
extracts. Various solvent system were used to ascertain the number of components. Six and seven phytochemicals were
recorded present in Yucca capensis and Ficus exasperate plants. The antibacterial studies revealed that Y. capensis extracts
have no antibacterial activities while it is only methanol extract of F. exasperate that showed slight antibacterial activities.
This study will validate the phytochemicals present in these plants and provide insight for more studies in their individual
and synergic activity in management of skin problem.
Introduction al., 2007). Skin disease is one of the five most medical
It is known that the lay people in rural areas of Nigeria consultation worldwide that affect people of all ages from
depend on medicinal plants for their primary health care. infant to the elderly and amount to approximately 34% of
However no survey has been done to document the all occupational diseases encountered (Abbasi et al., 2010).
phytochemicals present in medicinal plants used to treat Skin is an organ providing contact with the environment
various skin disorder. Nigeria is one of the developing and protecting the human body from unfavorable external
countries where about 80% of the population (Mukherjee, factors and plays a key role in protecting the body against
2002; Bodeker et al., 2005; Bandaranayake, 2006) use pathogens (Proksch et al., 2008; Madison, 2003), excessive
plant based remedies to prevent and cure several ailments. loss of moisture (Madison, 2003), as temperature
The prevention and cure of several ailments with medicinal regulation (insulator); synthesis, and storage of vitamin D
plants and herbal remedies varies from community to by action of ultraviolet (UV) and the protection of vitamin
community as various cultures have different herbs for B folates; absorption of oxygen and drugs (Grice et al.,
different diseases (Sharif and Bariki, 2006; Kubmarawa et 2009) and water resistance (Pappas, 2009). Skin disease is

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provoked by pathogens, noxious mechanical and chemical Scordato 1999; Blumenthal et al. 1998; Fleming 2000;
agents, and autoimmune responses. It is a complex process Bisset and Wichtl 2001).), essential oil (plant terpenes) are
during which the body repairs tissue damaged by forming most active component used in skin disease treatment
discolored and depigmented scar tissue and defends itself (Pittler and Ernst 1998; Swords and Hunter 1978), ajoene
against harmful stimuli (Dawid-Pać, 2013). (Ledezma, De Sousa, and Jorquera 1996), flavonoid was
There are a number of skin diseases that are disorders of found to possess highly protective effects against
exclusively (or predominantly) the superficial layers of the chemically induced skin tumor promotion in mice. (Lahiri-
skin. Skin diseases such as leprosy, endemic Chatterjee et al. 1999),bioflavonoids (proanthocyanidins)
treponematoses, or different varieties of filariasis and was beneficial by strengthening capillaries (Fleming 2000),
measles, chickenpox, and dengue fever with tissue sesquiterpene lactones such as helanalin, reduce
infections (e.g. erysipelas, cellulitis and abscess) are not inflammation by inhibiting the transcription factor nuclear
included as there is already an abundant literature on them factor κB (NF-κB) that controls the transcription of many
(WHO, 2005). genes, including cytokines such as interleukin and tumor
Skin disease such as rashes, viral infections, bacterial necrosis responsible for antigen presentation and activation
infections, fungal infections, parasitic infections, of cyclooxygenase 2 (Lyss et al. 1997). It is not
pigmentation disorders, tumors, cancers and other recommended for use on open wounds or broken skin, and
conditions (Wrinkles, rosacea, spider veins and varicose is included in Class 2d (McGuffin et al. 1997), mucilage
veins are among those conditions that cannot be neatly which is useful topically to soothe and act as an emollient
categorized) are common ailment that affects all ages from on skin and dries as a mild adhesive and can be used as an
the neonate to the elderly and cause harm in a number of herbal bandage for minor wounds (McGuffin et al. 1997;
ways (Marks and Miller, 2006). Peirce, Fargis, and Scordato 1999; Fleming 2000) saponins
Prevention of skin diseases is the best way of maintaining have softening and soothing effects and no adverse effects
healthy skin, on the other hand, many people may develop have been reported (McGuffin et al. 1997; Peirce, Fargis,
diseases that affect the skin including cancer, herpes, and and Scordato 1999).
cellulitis. There are many herbs that can be used in skin Fabaceae and Asparagaceae families are the most
diseases in order to relieve pain, burning, itching and even frequently used in Oghe traditional medicine for the
heal many of these conditions. Traditional medicine are the treatment of skin diseases (De Wet et al., 20 13).
best solution for all kinds of skin problems and are free MATERIALS AND METHOD
from any side effect of chemical drugs and also promise Collection of Plant Materials:
glowing and healthy skin, there was a call to return to The leaves of Yucca capensis (Ukpolodu onyeala) of
nature, natural remedies became a part of the green Asparagaceae family and Ficus exasperate (Awariwa) of
revolution, and there was a return to natural product Fabaceae family were collected from Oghe, Ezeagu Local
chemistry (Shenefelt, 2011). Government of Enugu State Nigeria. The herbarium
Below are some of the phytochemicals effective as specimens were authenticated by Prof. J.C Okafor of
remedies for your skin problems. Tannins are useful Department of Applied Biology and Biotechnology. The
astringent that soothe inflammation of the skin and mucous trees were photographed and the leaves were collected
membranes in disorders such as varicose veins and during the months of August, 2019.
hemorrhoids, act by coagulating the surface proteins of Extraction of Phytochemicals:
cells and exudates, thereby reducing permeability and The leaves of Y. capensis and F. exasperate were dried in
secretion. (McGuffin et al. 1997; Peirce, Fargis, and the laboratory at room temperature for a period of two

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weeks. The dried plant materials were powdered microorganism adopting standard disc diffusion method in
mechanically and kept in the air-tight containers. Thirty Nutrient agar medium. The concentrations of 200µg/disc
gram of the powder of each plant leaves was successively was taken for the extracts and also the control
extracted with n-hexane, ethyl acetate and methanol. The (erythromycin). The plates were incubated at 370C for 24h.
extracts were concentrated to a constant weight by The experiments were performedss in triplicates and the
evaporation. antibacterial activity was expressed as the mean diameters
Preliminary Phytochemical Investigations of clear zones of inhibition.
Qualitative phytochemical analysis of n-hexane, ethyl Minimum inhibitory concentration
acetate and methanol extracts of leaves of E. variegate and (MIC)Determination
F. exasperate were carried out by adopting the Minimum inhibition concentration was carried out using
characteristic colours produced by the reactions as broth dilution method as previously reported by Umer
observed and recorded as per the standard methods from (2013). The minimum concentration of the extracts of Y.
various sources. The bioactive compounds such as capensis and F. exasperate inhibiting growth of the three
alkaloids, flavonoids, glycosides, saponins, tannins, microorganisms’ were tested for MIC. Serial dilutions were
terpenoids, anthracene, resins and volatile oil were prepared from250 mg/ml of the extracts using DMSO to
screened by conducting three different tests for each make 250, 200, 100, 50, 25, 10, 5, 2 and 1 mg/ml solutions.
phytochemical to ascertain their presence (Krishnaveni. The test tubes were inoculated with (0.3mL) suspension of
and Ravi, 2014; Marandi and Britto, 2015; Kokate, 1994; the standard inoculated and incubated at 370C for 24h. The
Harborne, 1998; Khandewal, 2008; Trease and Evans, lowest concentration of the extracts which show no visible
1985; Peach and Tracey, 1956; Gibbs, 1974). growth of the broth was taken as the minimum inhibition
ANTIBACTERIAL SCREENING concentration.
Collection of micro-organism RESULTS AND DISCUSSION
Three clinical isolates (Staphylococcus aureus, The results of preliminary phytochemical analysis of
Streptococcus faecalis and Neisseria gonorrhea) were extracts of Y. capensis leaves revealed the presence of
obtained from Microbiology Laboratory, University of various phytoconstituents like of saponin, glycosides,
Nigeria Teaching Hospital, (UNTH) Ituku Ozalla, in triterpenoids, tannins, flavonoids and alkaloids. While all
Enugu Nigeria. The bacterial species were maintained at the extracts of F. exasperate leaves showed the presence of
370C on nutrient agar for the antibacterial studies of E. saponins, glycosides, tannins, triterpenoids, resins, volatile
variegate and F. exasperate. oil, flavonoids and alkaloids (Table 1).
Determination of antibacterial activity In the sensitivity report shown on Table 2, specimen such
The disc diffusion method was employed according to as urine, semen, high virginal swabs (HVS), urethral semen
Awodele et al., 2013. Stock solution (100mg/mL) of the (u/s), and sputum (cough) were cultured and organisms like
three extracts of leaves of E. variegate and F. exasperate Staphylococcus aureus, Streptococcus faecalis and
were prepared. Discs (6mm diameter) were cut from Neisseria gonorrhea were identified.
Whatman filter paper and sterilized at 2000C. The blank
sterile disc was placed on the inoculated agar surface and
impregnated with 20mL of stock solution using an
antibiotic disc of erythromycin (20µg) as control. The three
extracts of leaves of Y. capensis and F. exasperate were
tested for susceptibility against all the given

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Table 1: Results of phytochemical screening of Y. capensis and F. exasperate leaves extracts

Parameters Y. capensis Ficus exasperate
n- Ethyl- Methano n- Ethyl- Methano

Hexane acetate l Hexane acetate l
Saponin ┼ ┼ — — ┼ —
Saponin Glycoside — ┼ ┼ — — ┼
Hydrolysable ┼ ┼ — — — —
Tannins
Pseudo Tannin — — — — — —
Tannin (Using — ┼ ┼ — — ┼
Gelatin)
Digital Glycoside — — — — — —
Glycoside General — — ┼ — — —
Anthracene (Born — — — — — —
Tragger Test)
Resins — — — ┼ — —
Steroid and ┼ — — ┼ ┼ —
Triterpenoids
(Libemann
Buchard) Test
Volatile Oil Test — — — ┼ — —
Alkaloid — — ┼ — — —
a. Draggendoff Test
b. Wagner’s Test — — ┼ — — ┼
c. Mayer’s Test — — ┼ — — —
d. Kraint’s Test — ┼ ┼ — — ┼
Flavonoid Test — — — — — —
(a). Magnesium
ribbon test
(b). Alkaline test — — — — — —

(c). Lead Acetate — ┼ — — ┼ —
test
┼ = Positive, — = Negetive

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Table 1: Results of phytochemical screening of Y. capensis and F. exasperateleaves extracts

Parameters Y. capensis Ficus exasperate
n-Hexane Ethyl methanol n-Hexane Ethyl Methanol

acetate acetate
Saponin ┼ ┼ — — ┼ —
Saponin Glycoside — ┼ ┼ — — ┼
Hydrolysable Tannins ┼ ┼ — — — —
Pseudo Tannin — — — — — —
Tannin (Using — ┼ ┼ — — ┼
Gelatin)
Digital Glycoside — — — — — —
Glycoside General — — ┼ — — —
Anthracene (Born — — — — — —
Tragger Test)
Resins — — — ┼ — —
Steroid and ┼ — — ┼ ┼ —
Triterpenoi
ds (Libemann
Buchard) Test
Volatile Oil Test — — — ┼ — —
Alkaloid — — ┼ — — —
a. Draggendoff Test
b. Wagner’s Test — — ┼ — — ┼
c. Mayer’s Test — — ┼ — — —
d. Kraint’s Test — ┼ ┼ — — ┼
Flavonoid Test — — — — — —
(a). Magnesium
ribbon test
(b). Alkaline test — — — — — —

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Streptococcus faecalis and Neisseria gonorrhea (zone of

During the process of culture it was found that all the inhibition >10 mm). The ethyl acetate extract of F.
identified organism were sensitive to the standard drug exasperate showed weak activity against all the test
erythromycin. In this present study, antibacterial activities organisms. Hexane extract exhibited no or weak inhibitory
of Y. capensis and F. exasperate leaves extracts against effect against Staphylococcus aureus, Streptococcus
Staphylococcus aureus, Streptococcus faecalis and faecalis and Neisseria gonorrhea. The n-hexane, ethyl
Neisseria gonorrhea were assayed by agar well diffusion acetate and methanol extracts of Y. capensis exhibited no
method (Table 2). or weak activity against Staphylococcus aureus,
The antibacterial activities revealed that only the methanol Streptococcus faecalis and Neisseria gonorrhea (Table 2).
extract of F. exasperate had moderate activity against all
the organisms tested, mainly Staphylococcus aureus,
Table 2: Antibacterial activity of n-hexane, ethyl acetate and methanol extracts of Y. capensis and
F. exasperate leaves
Inhibition Y. capensis Ficus exasperate
(mm)
(mg/disc)
Test organism Test organism
Extract S. aureus S. faecalis N.gonorrhea S. aureus S. faecalis N. gonorrhea
n-Hexane
25 0±0 0±0 0±0 1 ± 0.3 0±0 1 ± 0.4
50 0±0 1 ± 0,3 0±0 1 ± 0.1 1 ± 0.4 2 ± 0.1
100 1 ± 0.1 1 ± 0.1 1 ± 0.1 2 ± 0.2 1 ± 0.3 2 ± 0.3
200 1 ± 0.2 1 ± 0.5 1 ± 0.2 3 ± 0.2 3 ± 0.1 2 ± 0.1
400 1 ± 0.1 1 ± 0.2 1 ± 0.2 3 ± 0.4 3 ± 0.1 3 ± 0.2
Ethyl
acetate
25 1 ± 0.4 1 ± 0.1 1 ± 0.2 2 ± 0.2 1 ± 0.4 2 ± 0.1
50 1 ± 0.2 1 ± 0.1 1 ± 0.1 4 ± 0.5 4 ± 0.3 3 ± 0.3
100 2 ± 0.5 1 ± 0.3 3 ± 0.1 5 ± 0.1 5 ± 0.1 3 ± 0.2
200 3 ± 0.2 2 ± 0.2 3 ± 0.7 7 ± 0.4 6 ± 0.3 5 ± 0.2
400 4 ± 0.5 3 ± 0.4 3 ± 0.3 7 ± 0.8 6 ± 0.2 6 ± 0.6
Methanol
25 2 ± 0.1 2 ± 1.3 1 ± 0.3 2 ± 0.1 4 ± 0.9 3 ± 0.4
50 2 ± 1.3 3 ± 2.0 4 ± 0.2 9 ± 1.5 10 ± o.5 8 ± 0.6
100 3 ± 1.4 5 ± 0.1 4 ± 0.4 11 ± 1.2 12 ± 0,7 12 ± 0.8
200 4 ± 0.2 6 ± 1.5 5 ± 1.2 12 ± 0.0 13 ± 0.1 14 ± 0.3
400 5 ± 0.8 6 ± 0.5 5 ± 0.7 14 ± 0.2 15 ± 1.1 16 ± i.1
Erythromycin 22 ± 0.5 25 ± 0.3 37 ± 0.7 22 ± 0.5 25 ± 0.3 37 ± 0.7
The methanol extract of F. exasperate leaves showed moderate antibacterial activity against the three
pathogens. MIC value of the methanol extract of F. exasperate leaves was presented (Table 3).

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Table 3: Minimum Inhibitory Concentration (MIC) of n-hexane, ethyl acetate and methanol
extracts of Y. capensis and F. exasperateleaves
Microorganism Y. capensisextracts F. exasperateextracts Standard drug
n-Hexane Ethyl Methanol n-Hexane Ethyl Methanol Erythromycin

acetate acetate
Staphylococcus Na 1 3 Na 2.5 10 125
aureus
Streptococcus Na 1.5 2 Na 6 12 63
faecalis
Neisseria Na 1 2 Na 5 11 46
gonorrhea
Na = not active and Neisseria gonorrhea. The antimicrobial activity of F.

exasperate were 125, 63 and 46 mg/ml concentration
The extended activities by both Y. capensis and F. (MIC) for Staphylococcus aureus, Streptococcus faecalis
exasperate extracts on microorganisms can be attributed to and Neisseria gonorrhea respectively. This work justifies
the presence of secondary metabolites such as terpenoids the further need to isolate and characterize the medicinally
(Scortichini and Rossi, 1991; Anyasorb et al., 2011; active compounds.
Esekhiagbe et al., 2009), flavonoids (Maji et al., 2010) and
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Pharmacology and Alternative Medicine Academic Journal

(PAMAJ); Vol.5, No.3, 2020;