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 American Journal of  
Food Science and Technology (AJFST)

Phytochemical Analysis on Aqueous Leaf  Extract of  Justicia carnea (Acanthaceae) and 
its Antibacterial Activity on Some Isolated Bacterial

O. Imohiosen1*  

Volume 2 Issue 1, Year 2023
ISSN: 2834-0086 (Online)

DOI: https://doi.org/10.54536/ajfst.v2i1.1564
https://journals.e-palli.com/home/index.php/ajfst

Article Information ABSTRACT

Received: April 13, 2023

Accepted: May 02, 2023

Published: May 07, 2023

The present study was aimed to investigate the bioactive constituents and antibacterial efficacy 
of  Justicia carnea leaf  aqueous extracts on some pathogenic organisms involved in causing 
infections in humans. The antimicrobial and medicinal properties was examined through 
phytochemical screening of  the plants constituents using standard qualitative methods 
and conducting bioassay on target bacteria of  medical importance such as Escherichia coli, 
Salmonella typhi, Staphylococcus aureus, Klebsiella pneumonia and Pseudomonas aeruginosa using the 
agar well diffusion method. The screening revealed that tannins, saponins and alkaloids is 
highly detected followed by flavonoids that is moderately detected whereas phlobatannins, 
phenol and volatile oils are present in low amount. The result shows that the leaf  extract 
was active against S. aureus, K. pneumonia, S. typhi, E. coli and P. aeruginosa measuring 
clear zones of  inhibition 26.0mm, 22.0mm, 20.0mm, 18.0mm and 18.0mm respectively. 
Hence, the extracts exhibit strong antibacterial activity and could be used as a preventive and 
curative measures to common diseases related to the test organisms.

Keywords

Analysis, Antimicrobial, J. 
carnea, Medicinal Properties, 
Phytochemical.

1 Department of  Science Laboratory Technology, Federal Polytechnic Bali, Taraba State, Nigeria
* Corresponding author’s e-mail: ojeaga2003@yahoo.com

INTRODUCTION
The use of  traditional medicine for healing purposes and 
to treat infections has been practiced since the origin 
of  mankind. It is getting ever more popular in Africa 
including Nigeria and has been part of  African culture 
in the recent years (Okonkwo, 2012). Serious clinical 
problems in the treatment of  infectious diseases has been 
created, since microorganisms have developed resistance 
to many antibiotics. Thus, Plethora of  studies has 
emerged towards using natural products as an alternative 
therapy (Kumari, et al., 2010). Medicinal plants have been 
the oldest traditional medicine considered important 
in treating several human ailments, including bacterial 
infections (Shyamapada & Manisha, 2011).

Justicia carnea Plant
The genus Justicia, named after the 18th-century Scottish 

botanist James Justice, belongs to the large family of  
Acanthaceae consisting of  about 600 species of  herbs 
and shrubs native to the tropics and subtropics. J. carnea 
is a flowering plant, widely distributed in various parts of  
Africa. In Nigeria, the shrubs of  J. carnea are grown around 
homesteads and act as fences, which are easy to grow and 
propagate from stem cuttings by pushing the stems 1 to 2 
inches into the soil (Corrêa & Alcântara,2012).

Figure 1: Picture of  Justicia Carnea 
Source: Fieldwork 2023.

Taxonomy/Classification of  Justicia carnea                                  
Kingdom Plantae
Phylum Tracheophyta
Class Magnoliopsida
Oder Lamiales
Family Acanthaceae
Genus Justicia L.
Species Justicia carnea Lindl.
Binomial name Justicia carnea(GBIF Secretariat, 2021; 

USDA PLANTS, 2007-2010).   

Chemical Composition of  Aqueous leaf  extract of  
Justicia carnea 
Some of  the phytochemicals investigated in the 
leaves include phenols, terpenoids, tannins, alkaloids, 
carbohydrates, flavonoids, saponins, phenols, reduced 
sugar and glycosides using standard procedures 
(Harborne,1998). Also present in the aqueous leaf  extract 
are vitamin A, B1, B12, B6, B9, B2, C and E containing the 
highest concentration of  vitamin C, and high concentration 
of  iron and calcium while magnesium, zinc and copper were 
present in low concentrations (Orjiakor et al., 2019). 
Hence, the aqueous leaf  extract of  Justicia carnea contains 
pharmacologically useful active principles elements. Thus 

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Am. J. Food. Sci. Technol. 2(1) 16-20, 2023

the aqueous leaf  extract of  the plant could play vital roles 
in health and disease.

Importance and Uses of  Justicia carnea 
Several species of  Justicia is one of  the many medicinal 
plants used in Nigeria and it has numerous ethno 
botanical uses. Traditionally, Justicia carnea is used in the 
management of  inflammation, gastrointestinal disorders, 
respiratory tract infection, fever, pain, diabetes, diarrhea, 
liver diseases, rheumatism and arthritis (Badami et al., 
2003; Corrêa & Alcântara, 2012). They also possess anti-
inflammatory, anti-allergic, anti-tumoral, anti-viral and 
analgesic activities (Radhika et al., 2013). 
Justicia carnea is a medicinal plant that is used conventionally 
as blood tonic from time immemorial in Nigeria. The 
resultant liquid after boiling is usually crimson red. Local 
consumers of  the leaves are mostly anaemic patients, 
women who want their blood replenished after the 
menstrual cycle and pregnant women. It is generally 
considered as an ornamental plant and it is widely 
distributed in various parts of  Africa. In Nigeria, it is 
grown around homesteads and acts as fences (Olufunke, 
2021).

LITERATURE REVIEW
The preliminary phytochemical analysis of  aqueous leaf  
extract of  J. carnea revealed that terpenoids, tannins, 
alkaloids, carbohydrates, flavonoids, saponins, phenols, 
reduced sugar and glycosides. Phenols and flavonoids were 
found in high concentrations which could be responsible 
for the anticancer and antioxidant activities exhibited 
by the plant (Orjiakor, Uroko, Njoku & Ezeanyika, 
2019). The presence of  terpenoids and carbohydrates 
in the aqueous leaf  extract of  the plant was against the 
findings of  Anigboro, Avwioroko, Ohwokevwo, Pessu & 
Tonukari (2019), who reported their absence in aqueous 
leaf  extract of  the same plant. Similarly, Onyeabo, Achi, 
Ekeleme-Egedigwe, Ebere & Okoro (2017) in their work 
also reported that phenols, alkaloids, tannins, flavonoids, 
terpenoids, saponins and steroids were present in the 
ethanol leaf  extract of  J. carnea, with high percentage of  
terpenoids, alkaloids and saponins. 
According to Igbinaduwa, Kabari & Chikwue (2019) 
alkaloids, flavonoids, tannins, phenols and saponins 
were present in the methanol leaf  extract of  J. carnea. In 
another research, flavonoids, alkaloids, phenols, tannins, 
carbohydrates, glycosides, gum, protein, fixed oils and 
fat were reported to be present in methanol leaf  extract 
of  J. carnea. While alkaloids, fixed oils were absent in the 
root extract of  J. carnea, sterols, gum was reported in both 
stem and root extracts (Sini, Prashy & George, 2018). The 
absence of  saponins in the methanol leaf  extract of  J. 
carnea was against the report of  Igbinaduwa et. al. (2019) 
who in their work reported the presence of  saponins. 

MATERIALS AND METHODS
Study Area  
The study was carried out in Federal Polytechnic Bali, Bali 

Local Government Area (LGA) of  Taraba State, Nigeria 
(see Fig.2). The Local Government Area lies between 
latitude 7046 N and 7054 N of  the equator and longitude 
10 030 E and 110 00 E of  the prime meridian (Topographic 
sheet, 1968). This falls within the dry guinea savannah 
with an estimated land area of  11,540 km2. It has some 
mountains like Gazabu, Dakka, Maihula, Bagoni, among 
others. Based on the 2006 National Population Census, 
Bali had a population of  about 211,024 persons (NPC, 
2006). 

Figure 2: Map of  Bali Local Government Area showing 
the study Area 
Source: Bureau  for Land and Survey Jalingo, Taraba State.

MATERIALS
Thermostated water bath,  Electronic balance 
(Model: 3003H), Hot plate (Model: ORL 2080), Oven 
(Thermcool), Desiccators, Cotton wool, Filter paper, 
Pestle and mortar, Glass wares (volumetric/conical flask, 
beakers, funnel), Justicia carnea leaves 

Reagents/ Chemicals
Methanol, Diethyl ether, Ammonia solution, Mayer’s 
regent, Wagner’s regent, Iron (III) Chloride, Hydrochloric 
acid, Ethanol, Acetic acid, Tetraoxosulphate (vi) (H2SO4) 
acid, Sodium hydroxide (NaOH) 

Preparation of  Laboratory Reagents for Phytochemical 
Analysis 
Wagner’s Reagent
2g iodine crystals and of  3g potassium of  iodide were 
dissolved in 100cm3 of  distilled water and properly   
stirred to homogenize into a solution.

Ferric Chloride Solution
5g of  ferric chloride (Iron III Chloride) was dissolved in 
50cm3 of  distilled water to form a homogenous solution 
of  ferric chloride. 

Dilute Solution of  Hydrochloric Acid
2cm3 of  hydrochloric acid was measured out using 

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Am. J. Food. Sci. Technol. 2(1) 16-20, 2023

measuring cylinder and poured into a 250cm3 beaker 
containing distilled water and stirred to form a 
homogenous dilute solution of  HCl. 

Collection and Identification of  Justicia carnea Leaves 
Fresh samples of  the leaves of  Justicia carnea were 
collected in the month of  March, 2023 from Daniya 
house residence, Bali, Taraba State, Nigeria. The plants 
was identified using pertinent taxonomic literature (GBIF 
Secretariat, 2021; USDA PLANTS, 2007-2010).  
The plant sample (leaves) were dried on the laboratory 
table of  Science Laboratory Technology Department, 
Federal Polytechnic Bali, after drying at room temperature 
the plant was then milled using pestle and mortar and the 
powder obtained were used for extraction.

Aqueous Extraction Preparation of  Justicia carnea 
Leaves  
100 gm of  fine powdered sample of  Justicia carnea was 
weighed and mixed with 500 mL of  distilled water in a 
round bottom flask. A reflux condenser was attached to 
the flask and inserted with a heating mantle. 
The mixture was reflux for one hour and filtered with 
Watman filter paper No. 1. The reflux was done twice 
again using new distilled water at each stage. The filtrate 
was heated to dryness by evaporation. Part of  the 
dry filtrate (extract) was used for phytochemical and 
antimicrobial screening. 

Procedures for Phytochemical Analysis
The phytochemical screening of  the plants constituents 
were assessed by using qualitative methods described by 
Trease and Evans, 2009. 

Alkaloid Test
Equal volumes of  the solvent extract (5cm3) and the 
Wagner’s reagent clean test tube and observed for some 
minutes. The presence of  alkaloid was indicated by a 
brown precipitate.

Phenol
2mL of  extract was added to 2mL of  ferric chloride 
solution (FeCl3), a deep bluish green solution is formed 
with presence of  phenols.

Phlobatannins
1cm3 of  hydrochloride acid and 1cm3 of  solvent extract 
were placed into a clean test tube and the tube was heated 
for about 10 minutes. Reddish green coloration indicates 
the presence of  phlobatannins.

Flavonoids
5cm3 of  the solvent extracts was placed into a test tube 
and few pieces of  magnesium chips were added, followed 
by concentrated hydrochloric acid in drops And then in 
excess. Reddish colouration indicates the presence of  
flavonoids.

Saponins (Froth test)
1g of  the sample was weighed into a conical flask in 10mL 
of  sterile distilled water was added and boiled for 5 min. 
The mixture was filtered and 2.5mL of  the filtrate was 
added to 10mL of  sterile distilled water in a test tube. The 
test tube was stopped and shaken vigorously for about 
30 second. It was then allowed to stand for half  an hour. 
Honeycomb froth indicated the presence of  saponins. 

Volatile oils
2.0 mL of  extract solution was shaken with 0.1 mL dilute 
sodium hydroxide and a small quantity of  dilute HCl. A 
white precipitate was formed with volatile oils. 

Tannin
3g of  the powdered sample was boiled in 50 mL distilled 
water for 3 minutes on a hot plate. The mixture was 
filtered and a portion of  the filtrate diluted with sterile 
distilled water in a ratio of  1:4 and 3 drop of  10% ferric 
chloride solution added. A blue or green colour indicates 
the presence of  tannins. 

Microorganisms
The microorganisms used are known to be potentially 
pathogenic to humans. They include Escherichia coli, 
Salmonella spp., Staphylococcus aureus, Klebsiella spp. and 
Pseudomonas aeruginosa. All the organisms’ strains were 
obtained as clinical isolates from Sancta Maria Clinic, 
Bali, Taraba State and Optimum Laboratory, Jimeta-Yola, 
Adamawa State, Nigeria.

Antimicrobial Susceptibility Testing
The antibacterial activity of  Justicia carnea leaf  aqueous 
extracts (100mg/ml) in vitro on the isolates was 
determined by the ager well diffusion method as described 
by (Olakunle et al., 2013 : Osho & Bello, 2010). This was 
done using pour plate method in which small colonies 
from each clinical isolates of  the test organisms were 
made into suspension with 1ml of  sterile distilled water 
in test tubes. 0.1ml of  each suspension was dispensed 
into sterile petri dishes after which melted and sterilized 
nutrient agar maintained at 450C was poured (15 aliquot) 
into the respective plates. The plates were allowed to set, 
four equidistant wells of  6mm in diameter were punched 
in each plate using a sterile cork borer. 0.2ml of  extract 
was introduced into each of  the wells,. A well filled with 
sterile water served as control and the plates were allowed 
to stay for15 minutes for pre-diffusion to take place 
followed by incubation for 24-48 hrs at 370C . The zones 
of  inhibition were measured with the use of  a metric rule 
and were recorded in millimeters.

RESULTS AND DISCUSSION
Table 1 revealed the presence of  some chemical/bioactive 
components in aqueous leaf  extract of  Justicia carnea. 
Tannins, saponins and alkaloids were highly detected 
followed by flavonoids that is moderately detected 

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Am. J. Food. Sci. Technol. 2(1) 16-20, 2023

Table 1: The Result of  Chemical Components of  Aqueous Leaf  Extract of  Justicia carnea
Phytochemicals    Test Aqueous Extract      
Alkaloids Wagner’s +++
Tannins  Ferric Chloride +++
Phlobatannins  Hydrochloric Acid + 
Phenol  Ferric Chloride +
Flavonoids  Magnesium Chips ++
Saponins Frothing +++
Volatile Oils Sodium Hydroxide +

+++= Highly Detected, ++= Moderately Detected, += Lowly Detected -= Not Detected

Table 2: Antibacterial activity of  Aqueous Leaf  Extract of  Justicia carnea against some bacteria isolates
Organisms (Bacteria) Dose (ml) Zone of  inhibition (mm)
Staphylococcus aureus 0.2 26
Salmonella typhi 0.2 20
Escherichia coli 0.2 18
Klebsiella pneumonia 0.2 22
Pseudomonas aeruginosa 0.2 18

whereas phlobatannins, phenol and volatile oils are present 
in low amount. These bioactive compounds have some 
therapeutic properties against several pathogens supporting 
its traditional use for curing disease. The presence of  these 
active agents confirms similar research conducted by 
(Orjiakor et al., 2019). Flavonoids have been acknowledged 
to posses potent antioxidant and free radical scavenging 
effect (Ganapth, et al., 2011) and have strong anticancer 
activity (Del-Rio, et al., 1997: Salah, et al., 1995). They are 
also documented to have anti-allergic, anti-inflammatory and 
anti-viral properties and help heal of  wounds and treat skin 
infections due to their ability to neutralize the acidity of  
wounds and inflammation (Okwu, 2004).
J.carnea extract also revealed to contain saponins. Saponin 
has the property of  precipitating and coagulating red 
blood cells. Other characteristics of  saponins include 
hemolytic activity, formation of  foams in aqueous 
solutions, cholesterol binding properties and bitterness 
(Okwu, 2004: Del-Rio, et al., 1997). More so, saponins 
have also been found to be an antibacterial substance 
on the cell wall of  most organisms (Harbron, 1998). It 
cause a reduction of  blood cholesterol by preventing its 
re-absorption. The presence of  alkaloid indicates that J.  
carnea extract can be used as basic medicinal agents for 
their antispasmodic, analgesic and bactericidal effects 
(Okwu & Okwu, 2004: Stray, 1998). 
The efficacy of  Aqueous Leaf  Extract of  J. carnea 
against different bacteria is shown in table 2. The sample 
showed effective antibacterial activity against both gram-
positive and gram-negative bacteria as indicated by their 
inhibition zones.  Staphylococcus aureus showed greater 
level of  susceptibility (26mm) than other tested bacteria. 
Followed by Klebsiella pneumonia and Salmonella typhi with 
level of  susceptibility (22mm and 20mm respectively). 
Escherichia coli and Pseudomonas aeruginosa showed the least 
zone of  inhibition (18mm). Hence, E. coli and P. aeruginosa 

were the most resistant isolates tested in this study as they 
both showed the least zones of  inhibition of  18mm.

CONCLUSION 
This present study showed that  J. carnea  leaf  is a good source 
of  important phytochemical or bioactive components 
which include tannins, saponins, alkaloids and flavonoids 
that are involved in both pharmacodynamics, bactericidal 
and bacteriostatic activity and these compounds are 
known to have some therapeutic properties against 
several pathogens which supports its traditional use for 
curing disease. They are vital sources of  supplementary 
diets of  several bioproducts essential for the biochemical 
functions in human, provide health benefits and are 
important for prevention illnesses. Therefore, justifies 
its usefulness in clinical and folklore practices against 
diseases caused by bacteria. Though, dosage remains a 
challenge.

Acknowledgements
This study was funded by the TETFund which is 
gratefully acknowledged. 

Conflict of  interest 
There is no conflict of  interest

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