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European Journal of Applied Sciences – Vol. 12, No. 4
Publication Date: August 25, 2024
DOI:10.14738/aivp.124.17375.
Bong, A. M., Benoit, N. M., Sohanang, N., Danielle, F. D., & Akah, N. V. (2024). Phytochemical Screening, Antimicrobial Properties
and Pharmaceutical Applications of Tapinanthus globiferus Leaf Extract. European Journal of Applied Sciences, Vol - 12(4). 353-
363.
Services for Science and Education – United Kingdom
Phytochemical Screening, Antimicrobial Properties and
Pharmaceutical Applications of Tapinanthus globiferus Leaf
Extract
Alang Michael Bong
Department of Applied Chemistry, ENSAI,
University of Ngaoundere, Cameroon
Ngassoum Martin Benoit
Department of Applied Chemistry, ENSAI,
University of Ngaoundere, Cameroon
Nodem Sohanang
Department of Food Engineering and Quality Control,
IUT, University of Ngaoundere, Cameroon
Fotio Defo Danielle
Department of Applied Chemistry, ENSAI,
University of Ngaoundere, Cameroon
Njong Vera Akah
Department of Chemical Engineering Catholic
University of Cameroon (CATUC), Bamenda
ABSTRACT
This study was carried out unveil the secondary metabolites of Tapinanthus
globiferus leaves used in traditional medicine through meticulous phytochemical
screening. The leaf extracts were obtained by methods of Soxhlet extraction and
maceration using water and ethanol as solvents. The leaf extracts were used to
carry out phytochemical screening, carry out some antimicrobial susceptibility
tests and indicate some industrial applications of the plant extracts. The extract
obtained by maceration in 95% ethanol gave a yield of 14.85% and extract tested
positive flavonoids and other polyphenolic compounds. For extended qualitative
analyses, the following secondary metabolites were identified; tannins, saponins,
anthraquinones, reducing sugars, alkaloids and terpenoids. The phenolic
compounds present give the extract antioxidant and antimicrobial properties. The
antimicrobial susceptibility tests were done on three bacterial species (E. coli with
27.78% inhibition, Staphylococcus aureus 28.33%; Streptococci 23.33%) and a
mold (C. albicans with a percentage inhibition of 26.67%). Results from
microbiological analysis showed that the plant extracts could inhibit the
proliferation of some pathogenic bacterial and fungal species. The minimum
inhibitory concentration (MIC) of the plant extract was 15.63 mg/mL for both E.
coli, S. aureus, and Streptococcus respectively while the MIC for C. albicans was 125
mg/mL. But the minimum bactericidal concentration (MBC) of the plant extract
against the bacteria was 125 mg/mL while the minimum fungicidal concentration
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Services for Science and Education – United Kingdom 354
European Journal of Applied Sciences (EJAS) Vol. 12, Issue 4, August-2024
(MFC) of the plant extract against C. albicans was 250 mg/mL. The outcome of this
research validates the use of this plant in traditional medicine and also validating
the plausible application of Tapinanthus globiferus in pharmaceutical and
cosmetic preparations for public health enhancement amongst others.
Keywords: Secondary metabolites, phytochemicals, Soxhlet extraction, pathogens,
public health, antimicrobial susceptibility.
INTRODUCTION
The dependence on medicinal plants in the rural areas of Cameroon and in other countries is
rising and in other countries for the treatment of various ailments due to the unavailability of
modern drugs in these areas coupled with the inability of the inhabitants of these areas to
afford modern healthcare. Most biological active natural products contain secondary
metabolites with a plethora of beneficial properties like antioxidant properties amongst
others [1]. In East Africa, more than 75 plants species from 34 families are used to cure more
than 59 ailments using traditional medicine and a whopping 80% of South Africans rely on
herbal remedies for their health care at various levels [2], [3], [4]. In Cameroon, more than
310 plant species are being used to treat various health ailments. Tapinanthus globiferus is a
parasitic plant that usually grows on the avocado tree (Persea americana) is being widely
used in Cameroon for the treatment of ailments like; typhoid, and malaria [5]. Mistletoes have
been identified as a remedy to more than 20 health predicaments [6]. Microorganisms are
increasingly resistant to modern drugs as reported by current researchers. This resistance
may be due to some inappropriate prescription and administration of drugs, ability of some
pathogens to undergo gene mutation, abusive use of antibiotics in food preservation and
other forms of drug’s misuse. Consequently, there is the need to seek recourse to alternative
sources of drugs with higher efficacies preferably from plant sources [7],[8]. Mistletoes of the
avocado tree have medicinal properties and are exploited as a form of traditional medicine in
Cameroon by a significant percentage of the population to resolve health predicaments like
malaria, typhoid inter alia. Medicinal plants are still being used as a therapeutic source, as
they play a vital role to resolve basic health needs of the population in developing countries
[9]. The phytochemical contents of plant species constitute the prime reason why they are
used as some form of medicine or drugs. Phytochemicals are bioactive compounds in plants
which have various health benefits to humans and other animal species [10]. Plants shall
continue to be a rich source of therapeutic agents and supply relevant industries. The
remarkable contribution of plants to the drug industry is made possible by the large number
of phytochemicals present in these plants [11]. This hemi-parasitic plant though used
traditionally as a medicinal source, little is documented about its phytochemical composition,
toxicology and beneficial chemistry. Recourse to traditional medicine is usually sort as it
depends on locally available plants which are easily accessible and cheap. Traditional
medicine however is limited as it is based on wisdom or knowledge handed down from
generation to generation without a scientific and clear methodology to prevent risks [12].
Mindful of the shortcomings of traditional medicine, poor knowledge of bioactive composition
of medicinal plants, this study was undertaken to screen the phytochemicals in Tapinanthus
globiferus, test their antimicrobial activities and recommend the use of this plant products for
pharmaceutical, cosmetic and related applications for public health amelioration activities.
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355
Bong, A. M., Benoit, N. M., Sohanang, N., Danielle, F. D., & Akah, N. V. (2024). Phytochemical Screening, Antimicrobial Properties and
Pharmaceutical Applications of Tapinanthus globiferus Leaf Extract. European Journal of Applied Sciences, Vol - 12(4). 353-363.
URL: http://dx.doi.org/10.14738/aivp.124.17375
MATERIALS AND METHODS
Materials
Tapinanthus globiferus plant leaves were harvested from an avocado tree (Persea americana)
at the country sides of Bamenda and Ngaoundere towns in Cameroon. The leaves of this plant
were identified by a botanist at the Agricultural Research Institute for Development (IRAD),
Bambili - Bamenda. The leaves were properly rinsed with clean tap water to remove dirt and
dust particles on them. Washed plant leaves were air-dried away from sunlight for 8 days, at
tropical room temperature. The dried leaves were then crushed using a moisture – free
blender. The plant leaf powder was sieved then packaged plastic bags and kept for use in the
research. Other materials included chemical reagents, solvents, glass ware, laboratory
equipment and microbial samples and accessories. Escherichia coli (E. Coli), Staphylococcus
aureus and Streptococcus were used as bacteria and Candida albicans were used as fungi
pathogens to determine their susceptibilities to the plant extract.
Methods
Extraction of Plant Leaf Phytochemicals (Bioactive Substances):
The chemical constituents of Tapinanthus globiferus leaves were extracted using the Soxhlet
apparatus and maceration. Distilled water was used as the solvent for Soxhlet extraction
while maceration was done using water and ethanol. The set-up for Soxhlet was done using
300 g of powder and extraction was allowed to continue until the solution in the siphon arm
of the Soxhlet extractor was colorless. For maceration, 700 g of leaf powder was mixed with 3
L of solvent at room temperature and corked in a plastic recipient. The set up was allowed to
stand for 4 days with occasional agitation. The solutions of the extract were separated using a
rotary evaporator. Then fine powder of the leave extracts was obtained and stored in dry
glass containers. Maceration was done following a modified method of Senhaji [13]. The yield
of secondary metabolites extracted was calculated as follows;
Yield = (
M1
Mo
) ×
100
1
....................................................... (02)
Where Mo = Mass of initial powder (g) and M1 = Mass of leaf extract (g)
Phytochemical Analyses of Tapinanthus globiferus Leaf Extract:
The various secondary metabolites in the plant extracts were qualitatively determined using
specific methods to screen for alkaloids, glycosides, saponins, steroids, phenols, tannins,
flavonoids, and terpenoids. For qualitative analysis, 10 g of powdered extract was dissolved in
100 mL of distilled water and solution used for some tests of bioactive substances following a
modified method of Trease and Evans [14].
Alkaloids were qualitatively determined using a modified method of Prashant [15]. In this
method, 2 mL of extract was taken in a test tube and 2 mL of 2M hydrochloric acid was added.
The mixture in the test tube was then treated with Wagner’s reagent. The formation of a
brownish / reddish precipitate indicates the presence of alkaloids in the crude extract.
Glycosides were screened using the Killer-Killiani Test, a modified procedure of Nidal [16].
Two milliliters of crude extract were measured into a test tube. 1 mL of glacial acetic acid