Arid Zone Journal of Engineering, Technology and Environment, March, 2018; Vol.

14(1):104-110
Copyright © Faculty of Engineering, University of Maiduguri, Maiduguri, Nigeria.
Print ISSN: 1596-2490, Electronic ISSN: 2545-5818, www.azojete.com.ng

QUALITATIVE PHYTOCHEMICAL SCREENING AND IN VITRO ANTIMICROBIAL

ACTIVITIES OF SOLANUM AMERICANUM MILL.

H. Usman,* V. Victor and I. Waziri

Department of Chemistry, University of Maiduguri, P.M.B. 1069, Maiduguri, Nigeria

*Corresponding author’s e-mail addresses: usmanhamidu@unimaid.edu.ng, husman321@yahoo.com,
Tel.: 0802 833 2246

Abstract
The use of plants as medicine is an ancient practice common to all societies especially the African societies and
particularly some part of Northern Nigeria. Solanum americanum leaf sample was collected at Kona Road Garden,
Bakin Kogi, Jalingo, Nigeria. The plant material was air-dried under shade and pulverized into coarse powder. After
extraction with reflux apparatus for four hours using 70% methanol, the air-dried concentrated extract was then
subjected to phytochemical evaluation and subsequent antimicrobial activities against some pathogenic
microorganisms: Bacillus subtilis, Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Klebsiella
pneumoniae, Pseudomonas aeruginosa and Candida albicans using standard methods of analyses. The
phytochemical results of the crude extract revealed the presence of alkaloids, anthraquinones, cardenolides,
saponins, tannins and flavonoids. The antimicrobial activity of the plant extract was assayed using disc diffusion
technique. The crude extract of S. americanum had the highest zone of inhibition of 15.00±0.00 mm against
Candida albicans while the least inhibition zone of 7.33±0.47 mm was found against Streptococcus pyogenes. Other
microorganism Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Klebsiella pneumoniae were found to
be resistant at all concentrations. These findings shows that S. americanum can only be effective against infections
related to Candida albicans and Streptococcus pyogenes. Hence, the use of S. americanum in traditional medical
practice for the treatment of some microbial-related infections.

Keywords: Solanum americanum, phytochemistry, Candida albicans, leaf, solanaceae

1. Introduction
Traditional medicine has been known for its mainstream practices not involving alternative
medicine. Traditional medicine is known as indigenous, folk medicine or home remedy practices
that develop over generation within various societies before the era of modern medicine. It is the
combination of the knowledge, skills, and practices base on the beliefs and experience
indigenous to different cultures used in the maintenance of wealth, prevention, diagnosis and
treatment of external and internal illness (Sofowora, 1993). A plant becomes medicinal when its
biological activity has been ethnomedically reported or scientifically established (Olaleye, 2007).
Medicinal plants are those that have similar properties as conventional pharmaceutical drugs
which have been identified and used from prehistoric times. These plants synthesize many
chemical compounds for biological functions; including defense against insects, fungi and
herbivorous mammals. Over 12,000 active compounds have been isolated and identified. These
phytochemicals work in a similar way as pharmaceutical drugs, so herbal medicines can be
beneficial and have harmful side effect just like the conventional drugs. Herbs are widely used to
treat diseases in non industrialized societies such as culinary herbs and spices. Angiosperms
(flowering plants) were the original source of most plant medicines (Dwinvedi and Smith, 2002).
In African ethno medicines, it is well known that traditional healers make use of a large variety
of herbs in the treatment of parasitic diseases including malaria and a wide proportion of herbal
remedies dispensed by traditional healers are widely believed by their clients to be effective

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(Bennett and Brown, 2000). Such plants can play an important role in drug discovery and their
studies are logical research strategies in search for new drugs (Kirby, 1997). Even though
pharmacological industries have produced a number of new antibiotics in the last three decades,
resistance to these drugs by microorganisms has increased. In general, bacteria have the genetic
ability to transmit and acquire resistance to drugs, which are utilized as therapeutic agents
(Gislene et al., 2000; Usman et al., 2017).

Solanum americanum (Solanum nigrum) has a long history of medicinal usage, dating back to
ancient times (Black Nightshade, 2017). This plant is an important ingredient in traditional
medicine for the treatment of dysentery, fever and helps in reducing gas formation in the
stomach and as well used as a strong analgesic and sedative with powerful narcotic properties; it
is also used topically for the treatment of harpes zoster, measles, itching, and eczema. The boiled
extracts of the leaves and berries are used to alleviate liver related ailments and to fight jaundice
(Black Nightshade, 2017). Solanum americanum belongs to the family Solanaceaa; a fast-
growing pioneer plant which helps to stabilize recently disturbed soil and thus makes it play an
important ecological role among the family of Solanaceae. S. americanum has received attention
as a vegetable worthy of domestication to supplement the nutritional demand of both rural and
central African communities. The plant is already a component of everyday diet and grows
rapidly, adapts well to a wide range of altitudes and soils; it has been reported to contain high
amount of protein, calories, fiber, calcium, iron and vitamin B and C (De Macuean, 2002).
Solanum americanum, commonly known as American black nightshade, Glossy nightshade,
popolo, popolohua; it is locally called “Gautan kadìì or Gautan kaajíí’’ in Hausa, “Oju
ologbo’’ in Yoruba and “Anya nwona’’ in Igbo language (Ibrahim and Nwude, 1980). The plant
has been used in the treatment of diarrhoea and dysentery in some parts of North-Eastern Nigeria
and thus the quest for scientific validation of this traditional claim.

2. Materials and Methods

2.1 Sample collection and identification
Fresh leaves of sample was hand-picked at Kona road garden, Bakin Kogi Jalingo, Taraba State
and was identified as Solanum americanum by a Taxonomist Mr. Emmanuel Chris of the
Department of Agriculture, Federal University Wukari, Nigeria; where a voucher number
CHM/16/005 was deposited. The sample was air-dried in the Laboratory at room temperature
and freed from twigs and dirt and then ground into coarse-powdered form with wooden mortar
and pestle.

2.2 Extraction procedure

Two hundred and fifty grams (250 g) of the powdered leaf sample of Solanum americanum was
extracted using 70% aqueous methanol by reflux technique for four (4) hours. The extract was
filtered using Whatman No.1. The filtrate was poured into evaporating dish and allowed to dry at
reduced temperature and pressure and then kept in a desiccator until use.
2.3 Phytochemical screening
Phytochemical Screening for the presence or absence of the secondary metabolites such as
alkaloids, anthraquinones, cardenolides, cardiac glycosides, flavonoids and saponins were
carried out according to the standard procedures as described by many researchers (Brain and
Tuner, 1975; Markham, 1982; Silva et al, 1998; Trease and Evans, 2002; Vishnoi, 1979).

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Usman et al.: Qualitative phytochemical screening and in vitro antimicrobial activities of solanum
americanum mill. AZOJETE 14(1):104-110 ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng

2.4 Antimicrobial susceptibility test

The antimicrobial susceptibility test was conducted using the agar plate disc diffusion technique
as described by Sidney et al. (1978); Vollekovà et al. (2001) with little modification by Usman et
al. (2005) . The tests were carried out using a stock concentration of 400 mg/ml prepared by
dissolving 4 g, 200 mg/ml was prepared by dissolving 2 g and 100 mg/ml prepared by dissolving
1 g of the crude extract differently into 10 ml of sterile distilled water. Working volumes was 0.2
ml each of the concentrations prepared and then dispensed into each of the 6 mm bored holes to
afford respectively 80, 40 and 20 mg/hole against the test microorganisms in triplicate. The
dilution ratio for Gram-positive bacteria and Gram-negative bacteria was 1:1000 and 1:5000
respectively using peptone water (Usman et al., 2005).

3. Results and Discussion

The phytochemical screening and antimicrobial activities were conducted on the crude leaf
extract of Solanum americanum. The results of the phytochemical evaluation shown on Table 1
revealed the presence of anthraquinones, cardenolides, alkaloids and flavonoids. The
susceptibility study of the extract shows dosages-dependent activity against some of test
microorganisms as presented on Table 2 below.
Table 1: Phytochemical analysis of crude extract of Solanum americanum leaf extracts
S/No. Phytochemical tests Inference
1 Test of alkaloids
Preliminary test for alkaloid +
Confirmatory test for alkaloid +
2 Test of free anthraquinones +
Test for combined anthraquinones +
3 Test of carbohydrates
Molisch`s test +
Test for combined reducing sugars +
Test of free reducing sugars +
4 Test for cardiac glycosides
Liebermann-Burchard’s test -
Salkowski’s test -
5 Test for cardenolides
Keller-Killiani’s test +
6 Test of saponins glycosides +
7 Test of tannins +
8 Test for flavonoids
Ferric chloride test +
Shinoda`s test +
Lead ethanoate test +
Sodium hydroxide test +
9 Test of Terpenoids -
Key: + = Present, - = Absent

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Table 2: Antimicrobial effects of the leaf extract of Solanum americanum

Concentration of the extract/Diameter of inhibition zones
(Mean±SEM)
Test microorganisms
20 mg/hole 40 mg/hole 80 mg/hole

Bacillus subtilis 0.00±0.00 0.00±0.00 0.00±0.00

Staphylococcus aureus 0.00±0.00 0.00±0.00 0.00±0.00
Streptococcus pyogenes 7.33±0.47 10.00±0.00 14.33±0.47
Escherichia coli 0.00±0.00 0.00±0.00 0.00±0.00
Klebsiella pneumoniae 0.00±0.00 0.00±0.00 0.00±0.00
Pseudomonas aeruginosa 0.00±0.00 0.00±0.00 0.00±0.00
Candida albicans 7.67±0.47 11.00±0.00 15.00± 0.00

In our continuous search for plant-drug with fewer side effects and potential activities against
microorganisms couple with the emergence of resistance to antimicrobial agents which does not
only result from poor quality drugs manufactured, patient non-compliance and irrational use of
antimicrobial agents, but also due to spontaneous mutations within the microbial populations;
(Nester et al., 2002; Denyer et al., 2004; Olajuyigbe et al., 2011) as well as selective pressure on
antibiotics (Buwa and Van staden, 2006). It is therefore, pertinent to screen this plant species
with the zeal to finding new potent drugs from medicinal plant. Moreover, it is well known fact
that infectious diseases account for high proportion of health problems, especially in the
developing countries like Nigeria. Table 1, shows the results of the qualitative phytochemical
screening of leaves extract of Solanum americanum plant. The result shows that anthraquinones,
cardenolides, alkaloids, flavonoids and carbohydrates were present in the crude extract of S.
americanum; whereas cardiac glycosides and terpenoids were absent. These classes of secondary
plant metabolites are known to show curative activity against several pathogens and therefore
could explain its use traditionally for the treatment of wide array of illness (Usman et al., 2005;
Usman and Osuji, 2007); Although, S. americanum have been reported to contain the presence of
toxic glycoalkaloids and there are warnings to be careful on the use of S. americanum as herbal
medicine and food (Mohy-ud-dint et al., 2010). Metabolites such as tannins, terpenoids,
alkaloids and flavonoids have been reported to have in vitro antimicrobial properties (Cowan,
1999; Dahanukar et al., 2000; Usman et al., 2013).

Table 2, shows the result obtained from the antimicrobial evaluation of the leaf extract of
Solanum americanum, the susceptibility pattern at concentration of 400 mg/ml (80 mg/hole) of
the extract of S. americanum crude leaf extract had highest inhibition zone for Candida albicans
species (15.00±0.00mm), followed by Streptococcus pyogenes (14.33±0.47 mm); while at
concentration of 200 mg/ml (40 mg/hole) it also showed highest inhibition zone for C. albicans
(11.00±0.00mm), followed by S. pyogenes (10.00 ± 0.00mm). The concentration of 100 mg/ml
(20 mg/hole) showed the lowest inhibition zone for C. albicans as (7.67±0.47 mm) followed by
S. pyogenes as (7.33±0.47mm). The resistant shown by B. subtilis was similar to that reported by
Valya et al. (2011), Similar research have been reported by Valya et al., 2011, who also worked
on the leaf extracts using four different solvents; this results at the working concentrations did
not corroborate with their work, who reported activities on E. Coli, S. aureus, P. aeruginosa at 5
mg per hole.

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americanum mill. AZOJETE 14(1):104-110 ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng

However, this work corroborates to their findings on C. albicans where inhibition was relatively
shown as well as dose-dependent pattern observed. Even though the extract showed a
concentration-dependent approach but lack a broad activities since its activity is centered on only
S. pyogenes and C. albicans with moderate activities. The zone of inhibition produce by most
antibiotic discs against some of the organisms were found to be more appreciative in relation to
those activities produced by most organisms under study though not statistically comparatively
to that presented by the extract. However, diameters of zones of inhibition of plant extractives
≥10 mm were considered active (Usman et al., 2005, Olajuyigbe et al., 2011).

4. Conclusions
In conclusion, this research work has laid credence to the traditional use of the leaf extract of
Solanum americanum as plant-drug against some superficial wounds infections, diarrhoea and
dysentery particularly against those whose causative agents are C. albicans and S. pyogenes.
These activities were primarily due to secondary metabolites found in the crude leaf extract.
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