Int.J.Curr.Microbiol.App.

Sci (2014) 3(12): 768-778

ISSN: 2319-7706 Volume 3 Number 12 (2014) pp. 768-778

http://www.ijcmas.com

Original Research Article

Phytochemical screening of the bioactive compounds in twenty (20)
Cameroonian medicinal plants

L.M. Ndam1*, A.M. Mih1, A.G.N. Fongod1, A.S. Tening2,

R.K. Tonjock3, J.E. Enang1 and Y. Fujii4
1
Department of Botany and Plant Physiology, University of Buea, P.O Box 63 Fako, South West
Region, Cameroon
2
Department of Chemistry, University of Buea, South West Region, Cameroon
3
Department of Biological Science, University of Bamenda, P.O Box 39 Bambili, North West
Region, Cameroon
4
Tokyo University of Agriculture and Technology, International Environmental and Agricultural
Sciences, Fuchu Campus, Saiwai-cho, Fuchu, Tokyo 183-8509, Japan
*Corresponding author

ABSTRACT

Preliminary screening of phytochemicals is a valuable step in the detection of

bioactive principles present in medicinal plants and may lead to novel
environmentally friendly bioherbicides and drug discovery. In the present study,
principal phytoconstituents of 20 Cameroonian medicinal plants were identified in
Keywords order to relate their presence with bioactivities of the plants. Screening of the plants
was performed using standard methods and resulted in the detection of the presence
Medicinal of tannins, flavonoids, phenolics, saponins, steroids, cardiac glycosides and
plants, alkaloids. Flavonoids were present in 18 of 20 plants while alkaloids were present
Allelopathy, in only four of the selected plants. It is evident from the study that Bridelia
Phytochemical, ferruginea, Justicia obliquifolia, and Morinda lucida registered the highest
Mount therapeutic efficacy, possessing majority of phytochemical classes of compounds
Bamboutos, while Spilanthes filicaulis recorded the lowest therapeutic potential due to absence
Wabane, of majority of phytoconstituents. These results validate the exploitation of the
Cameroon studied medicinal plants, according to high amounts of active principles and their
potential in medications, allelopathy, agroecosystems, dietary supplements or
cosmetics industries. Further studies are needed with these plants to evaluate their
allelopathic potentials, isolate, characterize and elucidate the structures of the
bioactive compounds responsible for their antimicrobial activity, allelopathic
activity, and other medicinal values.

Introduction

The tropical rainforest is home to a allopathic medicines used by humans

significant array of rich bio-resources and (Yorek et al.2008; Ndah et al., 2013).
accounts for 25 % of plant-extractable Medicinal plants are a group of species that

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accumulate different active principles, plants and more recently from micro-
useful in treating various human or animal organisms, with the animal kingdom
diseases. The long term use of herbs in contributing rather sparsely to the total (Fujii
medicine is a sure indication of their value et al., 1991; Duke et.al., 2000; Nazir et al.,
and usefulness in the future. In modern 2007). Over the last few decades, the
medicine, the importance of medicinal biological and pharmacological potentials of
plants is increasing (Iord chescu and organic substances from many indigenous
Dumitriu, 1988) with pharmaceutical and plants have been well understood. For
cosmetic industries increasingly using plant instance, phenolic compounds have been
resources from rural or unpolluted areas. associated with antimicrobial (Narayana et
al., 1999), anti-inflammatory, antiviral, and
Nature has been a source of medicinal cytotoxic activities (Chhabra et al., 1984).
agents for thousands of years and generally However, the bioactive constituents
produces many secondary metabolites which conferring these properties on many plant
constitute important leads for the species have also been implicated in
development of new environmentally allelopathy (Nazir et al., 2007).Allelopathy
friendly microbicides, pesticides, herbicides has been defined as the effect(s) of one plant
and many pharmaceutical drugs (Bobbarala on other plants through the release of
et al., 2009). Traditional societies in Africa chemical compounds in the environment
and elsewhere have always used plants to (Rice, 1984). Different plant parts, including
promote healing (Idu et al., 2005; flowers, leaves, leaf litter and leaf mulch,
Bussmann, 2006; Teferi et al., 2009) and stems, bark, roots, soil and soil leachates and
about 80 % of the world s population their derived compounds, can have
depends on the use of traditional medicine allelopathic activity that varies over growing
for health care (WHO, 1993). Therefore, seasons (Rice, 1984; Rizvi et al.,1999).
such plants should be investigated to better
understand their properties, safety and It has been indicated that phenolic acids are
efficacy (Doughari, 2008). the most commonly occurring natural
products noted for allelopathic activities
As science advanced, however, it became (Singh et al., 2003). Mungole et al. (2010)
possible to determine rigorously the active have also included alkaloids, coumarin,
components of these extracts through flavonoids, saponnins and volatile
painstaking and laborious chemical constituents of the essential oils as being
methods. This rational approach to the allelopathic agents. Generally, the presence
discovery of drugs inaugurated an era of of different phytochemicals in crude plant
bio-prospecting that is, raiding nature s extracts has been linked to the detrimental
storehouses of plant and microbiological effects of leachates, root exudates or
life. Bio-Prospecting literally involves decomposing residues of such plants on the
exploring the forests, diving in the oceans other vegetation or succeeding crops (Chung
and digging in the dirt to obtain et al., 2005; Mubashir and Wajaht, 2011).
environmental samples. The study of the Phytochemical analyses of several species of
compounds discovered by these methods has medicinal plants and allelopathic activities
become a major area of research in organic of the crude chemical compounds on crops
chemistry, biological science and has led to and plants have yielded positive results
the isolation and identification of thousands (Fujii et al., 2004). Of the different plant
of different structures, mostly extracted from families studied, Viles and Reese (1996)

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indicated that members of the Asteraceae interactions of medicinal species growing in

family have great potential for inhibitory association with traditional crops in the
activities. Thus, the increased interest in the Wabane municipality of the Mount
isolation and identification of the chemical Bamboutos Caldera of south western
compositions of organic products associated Cameroon. Most of these known medicinal
with biological activities with particular plants have not been studied empirically in
emphasis on germination, growth and yield detailed for the active chemical compounds.
of crops has stimulated research on plants It is now necessary to selectively screen
having both medicinal and allelopathic some of the medicinal plants for their
properties (Hegazy and Farrag, 2007). allelopathic or bioactive principles that
could be exploited for novel candidate
Cameroon is one of the richest floristic molecules for drug discovery and
regions of the world, well known for the bioherbicides for weed control. The aim of
different understory plant biomass and this work is to present for the first time, the
medicinal species (Sunderland et al., 2003; phytochemical screening of bioactive
Focho et al., 2009). It has diverse habitats compounds found in 20 of the most wide
with humid tropical forest covering 54 % of spread and used medicinal plants in the
the southern part of the country, mountain western flank (Wabane Municipality) of the
forest and savannah in the highlands and Mount Bamboutos Caldera of South
sub-Sahelian savannah and near desert in the Western Cameroon. The obtained result is a
far North (Sunderland et al., 2003). These key aspect in making recommendations
diverse habitats harbour more than 9,000 concerning the cultivation of certain species,
species of plants, 160 species of which are whose active principles can be valued as
endemic. The majority of the endemic taxa phytoterapeutical products, food
are concentrated around Mount Cameroon supplements, cosmetics and agroecosystem
and other highland areas such as the Mount chemical products.
Bamboutos Caldera in south western
Cameroon. Materials and Methods

The caldera is endowed with different plant Plant material

species valuable for health care needs of the
larger populations and fodder for the An inventory comprising about 200 species
livestock (Ayonghe and Ntasin, 2008; of medicinal plants identified in the Mount
Harvey et al., 2010; Fonge et al., 2013). Bamboutos Caldera was made by a research
Like many other developing nations, where team lead by the first author. From these, a
several classes of plant secondary total of 20 most commonly used species in
metabolites have been implicated in primary health care were selected and
allelopathy and biological control, similar analyzed in terms of their active principles.
observations have also been documented in The plants were harvested in different
Cameroon (Cho-Ngwa, 2010; Fongod, localities of the western flank of the Mount
2004). Worldwide, utilization of the Bamboutos Caldera of Cameroon and
allelopathic characteristics of both plants identified and authenticated at the Limbe
and crop cultivars for the control or Botanic Garden (LBG) by the botanist, Mr.
management of other vegetation in crop Litonga Ndive Elias with voucher specimens
based agroecosystems is common. However, deposited. Detailed information on each
very little is known on the allelopathic medicinal plant is given in Table 1.

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Preparation of extracts was dissolved in methanol and 2 mL of

concentrated hydrochloric acid added. A
Fresh plant materials were collected for each spatula full of magnesium turnings was
plant and dried at room temperature in an added and the mixture observed for
aerated laboratory for three weeks. The effervescence. A brick red colouration
dried materials were ground using a mill observed indicated the presence flavonoids.
with 2 mm sieve attached to it to yield a fine
powder. One hundred grams of each powder Test for steroids
was weighed and macerated three times in (Lieberman-Burchard test)
1000 mL of acetone for 48 hours. The
mixture was filtered using Whatman filter About one half gram (0.5 g) of the crude
paper No. 1 and the filtrate concentrated extract was dissolved in 0.5mL
under reduced pressure by rotary dichloromethane to give a dilute solution
evaporation (BUCHI Rotavapor R-200, and then 0.5 mL of acetic anhydride added,
Switzerland) at appropriate temperature. followed by three drops of concentrated
Residual solvent was removed by drying in sulphuric acid. A blue-green colouration
air at room temperature (23 25 °C) and the indicated the presence of steroids.
extract weighed and stored at -20 °C until
used. An aliquot of each crude extract Test for tannins (Ferric chloride test)
obtained was used for phytochemical tests
while the remaining fraction was kept for One half gram (0.5 g) of the crude extract
further studies. was dissolved and added to a tube
containing 20 mL of boiling distilled water
Qualitative tests: phytochemical and then boiled for an hour. A few drops of
screening ferric chloride was added and allowed to
stand for proper colour development. A
The concentrated residues from the acetone blue-black colouration indicated the
extracts were used to detect the secondary presence of tannins.
plant metabolites including alkaloids,
flavanoids, steroids, saponins, glycosides, Test for Alkaloids (Dragendorf s test)
phenolics and tannins using standard
methods with some modifications (Trease, The sample was dissolved in
1989; Christen, 2000; Young and Woodside, dichloromethane and then spotted on a thin
2001; MacNee, 2005) layer chromatographic plate which was
developed in 20 % hexane in ethylacetate.
Test for saponins (Frothing test) The presence of alkaloids in the developed
chromatogram was detected by spraying
Saponins were tested by dissolving one half with freshly prepared Dragendorf s reagent
gram (0.5 g) of the crude extract in a test in a fume chamber. A positive reaction on
tube containing 3 mL of hot distilled water the chromatogram indicated by an orange or
and then the mixture was shaken vigorously darker coloured spot against a yellow
for one minute and persistent foaming background is confirmatory evidence that
observed indicated the presence of saponins. the plant extract contained alkaloids.

Test for flavonoids (Cyanidine test) Test for cardiac glycosides

One half gram (0.5 g) of the crude extract An extract of the plant was added to 2 mL
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of glacial acetic acid plus one drop of ferric Test for phenolics
chloride. The set up was underplayed with 1
ml of concentrated sulphuric acid. There To 1 mL of the plant extract, one drop of 5
was the appearance of violet and brownish % FeCl3 (w/v) was added. Formation of
rings below the interface, followed by the greenish precipitate indicated the presence
formation of a greenish ring in the acetic of phenolics.
acid layer which indicated the presence of
cardiac glycosides.

Table.1 List of 20 Cameroonian medicinal plants used in phytochemical tests

S.No Plant Name Familly Life Part

form used
1 Ageratum sp. Asteraceae Herb Whole
2 Eremomastax speciosa Benth Acanthaceae Herb Leaf
3 Justicia obliquifolia Hochst Acanthaceae Herb Leaf
4 Ageratum conyzoides L. Asteraceae Herb Leaf
5 Emelia coccinea (Sims) G. Don Asteraceae Herb Whole
6 Spilanthes filicaulis (Schum. And Thonn) C.D. Asteraceae Herb Whole
Ada
7 Aframomum melegueta K. Schum Zingiberaceae herb Seed
8 Piper umbellatum L. Piperaceae Herb Leaf
9 Bryophyllum pinnatum (Lam.) Oken Crassulaceae Herb Leaf
10 Laportea ovalifolia (Schum & Thonn.) Chev. Urticaceae Herb Whole
11 Harungana madagascariensis Lam. & Poir. Hypericaceae Tree Bark
12 Impatiens macroptera Hook F. Rank Balsaminaceae Herb Leaf
13 Melanthera scandens (schumach and Asteraceae herb Leaf
Thonn.)Robety
14 Echinops gingateus Var. lelyi C.D. Adams Asteraceae Shrub Root
15 Vernonia hymenolepis A.Rich Asteraceae Shrub Leave
16 Vernonia comferta Benth Asteraceae Shrub Leaf
17 Momordica foetida Schum Cucurbitaceae Climber Leaf
18 Bridelia ferruginea Benth. Euphorbiaceae Tree Bark
19 Morinda lucida Benth Rubiaceae Tree Bark
20 Commelina benghalensis Linn. Commelinaceae Herb Whole

Results and Discussions

After performing the analysis of bioactive tannins, saponins, flavonoids, were detected
compounds of the studied medicinal plants as present in the medicinal plants in
extracts, results obtained are as shown in different proportions and classes. The results
Table 2. The phytochemicals, steroids, from the phytochemical screening of the
alkaloids, phenolics, Cardiac glycosides, studied medicinal plants extracts have
shown that flavonoids are found in eighteen
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of the twenty plants, with Eremomastax 2008.) and tannins exhibit antioxidant,
speciosa, Justicia obliquifolia, Momordica antimicrobial and antiviral effects (Sayyah
foetida and Bridelia ferruginea extracts and Hadidi, 2004).
being very rich in these compounds.
The plant extracts were also revealed to
Steroids are found in most of the plant contain steroids, which are known to
extracts, except for those obtained from produce an inhibitory effect on
Aframomum melegueta, Harungana inflammation (Savithramma and Linga,
madagascariensis and Echinops giganteus. 2011) and alkaloids that have been reported
In contrast, the extract from Justicia to exert analgesic, antispasmodic and
obliquifolia and Emelia coccinea were very antibacterial activities (Nyarko and Addy,
rich in steroids. The highest contents of 1990). The phytochemical screening results
alkaloids were found in Bridelia ferruginea of the extracts are consistent with the results
followed Piper umbellatum while seventeen reported by Alghazeer and El-Saltani
plant extracts did not contain this type of (2012), where authors mentioned the
compounds. Saponins were present in six presence of tannins, alkaloids, saponin and
studied plants, with Bridelia ferruginea terpenoids in screened medicinal plants.
habouring the highest content. Piper Phenolic acids are the most commonly
umbellatum and Morinda lucida are occurring natural products noted for
moderately rich in tannins, while sixteen of allelopathic activities (Singh et al., 2003).
the plant extracts did not contain these
compounds. Mungole et al. (2010) have also included
alkaloids, coumarin, flavonoids, saponnins
Cardiac glycosides were found highly and volatile constituents of the essential oils
expressed in the analyzed extracts of as being allelopathic agents. Generally, the
Justicia obliquifolia, Morinda lucida and presence of different phytochemicals in
Piper umbellatum. Phenolics are present in crude plant extracts has been linked to the
great quantities in Aframomum melegueta, detrimental effects of leachates, root
Harugana madagascariensis and Bridelia exudates or decomposing residues of such
ferruginea. Analyzing the results further, it plants on the other vegetation or succeeding
can be observed that the studied medicinal crops (Chung et al., 2005; Mubashir and
plants containing the largest number of Wajaht, 2011). It is difficult to compare the
bioactive compounds were Bridelia data with the literature because several
ferruginea, Justicia obliquifolia, and variables influence the results.
Morinda lucida while Spilanthes filicaulis
registered the lowest presence of According to some authors, the quantity and
phytochemicals. the composition of bioactive compounds
present in plants are influenced by the
The studied bioactive compounds have a genotype, extraction procedure, geographic
broad range of biological activities. For and climatic conditions, and the growth
example, phytochemicals such as saponins phase of the plants (Ciulei and Istodor,
have anti-inflammatory effects (Vinha and 1995; Trease and Evans, 2002). Plant cells
Soares, 2012), hemolytic activity, and produce two types of metabolites. Primary
cholesterol binding properties (Nyarko and metabolites are involved directly in growth
Addy, 1990). Glycosides are known to lower and metabolism (carbohydrates, lipids and
blood pressure (Marinkovic and Vitale, proteins).

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Table.2 Phytochemical constituents of twenty (20) Cameroonian medicinal plants

Plants Phytochemical constituents

Steroids Alkaloids Cardiac Phenolics Tannins Flavonoids Saponins
glycosides
Ageratum sp. + - - + - + -
Eremomastax speciosa (Benth) + - + - - +++ +
Justicia obliquifolia (Hochst) +++ - +++ - - +++ +
Ageratum conyzoides(Linn.) + - - - ++
(Cufod.) Dandy
Emelia coccinea (Sims) G. Don +++ - - + - ++
Spilanthes filicaulis (Schum. + - - - - + -
And Thonn) (C.D. Ada)
Aframomum melegueta (k. - + - +++ + + +
Schum)
Laportea ovalifolia (Schum & ++ + - - ++ -
Thonn.) (Chev.)
Harungana madagascariensis - + + +++ - + ++
(Lam. & Poir.)
Impatiens macroptera Hook F. + - + - - ++ -
Rank
Melanthera scandens (schumach + - + - - - -
and Thonn.)Robety
Echinops gingateus Var.lely D.C - - + + - + -
Vernonia hymenolepis A.Rich ++ - ++ ++ - ++ ++
Vernonia comferta Benth + - - - - + -
Momordica foetida Schum ++ - ++ + - +++ +
Bridelia ferruginea Benth. + +++ ++ +++ + +++ +++
Morinda lucida Benth ++ - +++ + ++ ++ -
Commelina bengalensis Linn. + - - - - ++ -
Piper umbellatum L. + ++ +++ + ++ - -
Bryophyllum pinnatum (Lam.) ++ - - + - + -
Oken

Legend: +=Low concentration, ++ = Moderate concentration, +++ = High concentration, - = Absent

Most natural products are compounds flavonoids, lignins, tannins, etc.) (Pal,
derived from primary metabolites such as 2007). These secondary metabolites are
amino acids, carbohydrates and fatty acids the major source of pharmaceuticals, food
and are generally categorized as secondary additives, fragrances and pesticides, and
metabolites. Secondary metabolites are herbicides (Okwu, 2005; Ramawat and
considered products of primary Dass, 2009; Ramu and Mohan, 2012).
metabolism and are generally not involved Similar analyses have been conducted in
in metabolic activity (alkaloids, phenolics, areas that have a long tradition in the
essential oils and terpenes, sterols, cultivation and utilization of medicinal
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