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Phytochemical Screening and Antimicrobial 
Activity of Terminalia catappa (L.) seed oil 
obtained from Anyigba, Kogi State, Nigeria, 
on Some Selected Clinical Isolates 

 P.R.O. Edogbanya, M.O. Suleiman, J.B. Olorunmola, R.F. 
Aminu and I.J. Oijagbe 

All Res. J. Biol., 2020, 11, 1-5 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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ARTICLE 



                                                          Issue 1, Vol 11, 2020, 1-5 

Phytochemical Screening and Antimicrobial Activity of 
Terminalia catappa (L.) seed oil obtained from Anyigba, Kogi 

State, Nigeria, on Some Selected Clinical Isolates 
P.R.O. Edogbanya *1, M.O. Suleiman1, J.B. Olorunmola1, R.F. Aminu2 and I.J. Oijagbe 3 

 
1Department of Plant Science and Biotechnology, Kogi State University, Anyigba, Nigeria. 
2Department of Microbiology, Kogi State University, Anyigba, Nigeria. 
3Department of Biology, Ahmadu Bello University, Zaria, Nigeria. 
*Corresponding Author: ocholiedogbanya@gmail.com +2348138381212 
 

Grafical Abstract    

 

 Abstract:  

This research was carried out in a bid to seek for alternative ways of dealing with strains of microorganisms that 
have become resistant to conventional antibiotics. The study was carried out to evaluate the antimicrobial 
efficacy of Terminalia catappa seed oil obtained from Anyigba, Kogi State Nigeria, on some selected clinical 
isolates of selected microorganisms (Aspergillus niger, Staphylocuccos aureus, Escherichia coli). The oil was 
extracted from the seeds using the soxhlet extraction method with n-hexane as the solvent. The Mueller Hinton 
(well diffusion method) was used to test the susceptibility of the strains of the microorganisms to the oil, using 
Ciproflaxin as standard positive controls, alongside Tween and DMSO as negative control. Experiments were 
carried out in duplicates. The results obtained revealed that Terminalia catappa oil was unable to create any 
inhibition zones on the clinical isolates. From this research, it can be concluded that Terminalia catappa oil had 
no Antibacterial activity. 

Keywords: Antimicrobial, Terminalia catappa, Essential oils, seed 

 

Introduction 

Although many different antibacterial agents are available in 
the field of medicine, many of these agents are increasingly 
being incapacitated by the microorganisms through the 
evolution of different mechanisms that amount to resistance 
to these drugs.1 Presently in the developing countries, 

synthetic drugs are not only expensive and inadequate for the 
treatment of diseases but also are often with adulterations and 
side effects.2 There is therefore a continuous and urgent need 
to discover new antimicrobial components with diverse 
chemical structures and novel mechanisms of actions because 
of the increase in the incidence of new and re-emerging 
infectious diseases3 to replace those that have lost their 

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All Res. J. Biol, 2020, 11, 1-5 

  

efficacy. Research has, however, shown that many herbs 
possess varying degree of antimicrobial activities. Kaufman 
et al. (1999)4 had reported that more than 25% of the 
prescribed drugs contained at least one active ingredient of 
plant origin and about 80% of world population relies on 
traditional medicine for significant part of their primary 
health care needs.  

Belonging to the family Combretaceae, Terminalia catappa 
Linn. is naturally occurring and widespread in the subtropical 
and tropical zones of the Indian and Pacific Oceans, and is 
planted extensively in many countries as an ornamental tree.5 
The type of oil that is obtained from Terminalia catappa 
seeds is safe for consumption, in addition Oliveria et al., 
(2000)6 indicated that the oil content of Terminalia catappa 
(583.0 g/kg dry matter) is comparable to that of other oil 
seeds such as peanut, rapeseed, and sunflower.  
Termianalia catappa is used primarily as an ornamental 
shade, and salt-tolerant street tree, but the leaves provide 
food for the Tasar silkworm, and the seeds are edible like 
almonds with similar oils. On the Malay Peninsula and 
through the Canary islands this tree is known as the tropical 
almond. Termianalia catappa has been claimed to have 
therapeutic effects for liver related diseases.7 It is attributed 
with cholagogue action. In India, it is used as cardiac 
stimulant. Its leaves are widely used as a folk medicine in 
Southeast Asia for the treatment of dermatosis and hepatitis8. 
More and more pharmacological studies have reported that 
the extract of Termianalia catappa leaves and fruits have 
anticancer by (Fan et al., 2004)9, antioxidant by (Zhai et al., 
2001)10, anti-HIV reverse transcriptase, anti-inflammatory, 
and  anti-diabetic effects as stated by (Xu et al., 2000)11 and 
hepato-protective activities as stated by (Nagappa et al., 
2003)12 but the effective components and related mechanisms 
remain unknown.  
Numerous studies have been conducted with the extracts of 
various plants, screening antimicrobial activity as well as for 
the discovery of new antimicrobial compounds13,14,15,16. The 
efforts of scientists in establishing plants with promising 
antimicrobial property is yielding fruitful results as a number 
of plants with high antimicrobial property have been 
elucidated. This work was carried out to determine the 
phytochemical components as well as evaluate the 
antimicrobial activity of Terminalia catappa seed oil. 
 
Methodology  
 
The essential oil used in this study was extracted from nuts of 
Indian almond. Ripe fresh seeds were collected directly from 
the tree plant and dried in an oven for 5 days to reduce 
moisture content and enable easy breaking to get reasonable 
amount of the nuts. The seeds were cracked open with a nut 
cracker and the nuts are pounded using mortar and pestle into 
a smooth powder to increase surface area so that the solvent 
can permeate properly into it to extract the oil. 
 
Extraction of essential oil by soxhlet method 
 
The extraction of essential oil was done using n-hexane in 
electro-thermal soxhlet extractor (Gallenkamp, England). 30g 
of powdered seeds was weighed and put into the thimbles of 

the soxhlet extractor, the apparatus was mounted and allowed 
to run for 6 hours, after which the mixture of essential oil and 
n-hexane was collected in a beaker and evaporated over a hot 
water bath to collect the oil as residue. The method was 
repeated thrice to get sufficient amount of oil.17 
 
 
Determination of relative density of oil 
 
Two density bottles were washed and dried in an oven and 
allowed to cool in a desiccator. Each of the bottles was then 
weighed empty and the weights recorded. One was filled 
with water and the other with Terminalia catappa seed oil. 
Both was weighed again and the weight of the water and oil 
were determined by the difference in weight.18 

 

 
Where: 
Weight of empty bottle = W1 
Weight of empty bottle + oil = W2 
Weight of empty bottle + water = W3 
 
Determination of the phytochemical constituents of oil 
 
Chemical tests is carried out on the seeds oil of Terminalia 
catappa using standard procedures to identify the 
constituents as described by (sofowora, 1982)19, (Trease and  
Evans, 1989)20 and (Harborne,1973)21.   
Alkaloids: About 0.5ml of the extracts is warmed with 2% of 
the H2SO4 for two minutes. It was filtered and a few drops of 
Dragondoff reagent is added. Orange red precipitate indicates 
the presence of alkaloids. 
Tannins: Small quantity of extracts is mixed using water and 
heated on water bath and filtered. Few drops of ferric 
chloride is added to filtrate. Dark green solutions indicates 
the presence of tannins.  
Glycosides: Extracts is hydrolyzed using HCL and 
neutralized with NaOH solution. Few drops of Fehling’s 
solution is added. Red precipitate indicates the presence of 
glycosides. 
Saponins: About 0.5ml of the extract is shaken with 5ml of 
distilled water and then heated to boil. Frothing (appearance 
of creamy miss of small bubbles) shows the presence of 
saponins.  
Flavonoids: Extract of about 0.5ml is dissolved in diluted 
NaOH and HCL added. A yellow solution that turns 
colourless indicates the presence of flavonoids. 
Total phenol: Extract of about 0.5ml is dissolved in Dennis 
solution. A red colour precipitate indicates the presence of 
phenol.  
 

Culture of Clinical Isolates 

Clinical isolates  (Aspergillus niger, Staphylocuccos aureus, 
Escherichia coli) were obtained from the Department of 
Microbiology, Faculty of Natural Sciences, Kogi State 

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University, Anyigba. Overnight cultures were obtained using 
10ml Nutrient agar and used for the study. 

Well Diffusion Assays 
 
Antimicrobial activity testing was carried out by using agar 
diffusion method. Three dilutions (30, 60 and 90%v/v) of 
Terminalia catappa seed oil were made using 0.005%v/v 
Tween 20 with 0.005%v/v Dimethyl sulfoxide (DMSO) in 
the ratio 1:1 as a co-diluent. Mueller-Hinton sterile agar 
plates were flooded with 2ml indicator microbial stains 
(Aspergillus niger, Staphylocuccos aureus, Escherichia coli), 
and the excess drained. A cork borer was flamed and used to 
bore five wells which contained  0.005% tween and 0.005% 
DMSO (1:1) (negative control); 30%, 60%, and 90% v/v of 
oil (treatments), and  0.5mg/L Ciprofloxacin (positive 
control) and allowed to stay at 37ᵒC for 3 hours. The zones of 
growth inhibition around the disks were measured using 
venier caliper and a meter rule after 18 to 24 hours of the 
incubation at 37ᵒC for bacteria and 48 to 96 hours for fungi at 
28ᵒC. The experiments were done in duplicates. 

Statistical Analysis 
One way Analysis of Variance (ANOVA) was used to 
compare the mean inhibition zones of the different treatment 
groups and Duncan Multiple Range Test (DMRT) was used 
to separate means where significant. P < 0.05 was considered 
significant.  
 
Results  
Results revealed that essential oil of Termianalia catappa 
seed contained tannins, flavonoids and glycosides (Table 1) 
 
Table 1: Phytochemical Components of Oil Extract 

Parameter Test/Reagent Observation Inference 

Alkaloids Dragondorff No colour change (-) 

Tannins Ferric chloride Dark green colour (+) 

Saponins Frothing test No frothing (-) 

Flavonoids NaOH-HCL test Slight yellow 
colour 

(+) 

Glycosides Fehlings A and B 
test 

Slightly red 
colour 

(+) 

Phenols Dennis test No colour change (-) 

Key: + Present    -  Absent 
 
Relative Density of the Oil 
The relative density of Terminalia catappa oil obtained was 
0.875 (Table 2) 
 
 
 

 
Table 2: Relative Density of Terminalia catappa Oil 
Sample Weight of 

Empty 
Density 
Bottle 

Weight of 
Empty Bottle 

× 

Sample 

Mass of 
Substance 

Density g/ml 

Oil 9.55 18.35 8.80 0.875 

Water 9.55 19.61 10.06  
 
 
Effect of the Extracts on Isolates 
 
Results showed that the extract (Termianalia catappa seed 
oil), at 30% (A) , 60% (B) , and 90% (C) v/v had no 
significant effect on the test organisms, the three organisms 
being Escherichia coli, Staphylococcus aureus , and  
Aspergillus niger respectively. The negative control (NC), 
like the extract, had no effect as well on all the test 
organisms. The positive control (PC) however had effects on 
E.coli, S. aureus, A. niger with inbition zones of 12mm, 
17mm and 22.5mm respectively (Table 3; Fig 1 -3). 
 
Table 3: Antimicrobial activity of Terminalia catappa seed oil 
extract on a few selected pathogenic organisms 

 

Pictorial Presentation of the Agar Plates 
 
 

                      
 

Figure 1: Effect of Terminalia catappa seed oil on Escherichia coli 
: NC = Negative Control (0.5% DMSO and 0.5% Tween 20) ; A = 
30% oil extract ; B = 60% oil extract ; C = 90% oilextract; PC = 
Positive Control (0.5mg/L ciprofloxacin)  

 

Test 
Organism 

Inhibition zone(mm) 

NC A B C PC 

E. coli - - - - 12.0 ± 2.0 

S. aureus - - - - 17.0 ± 3.0 

A. niger - - - - 22.5 ± 2.5 

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Figure 2: Effect of Terminalia catappa seed oil on Staphylococcus 
aureus: NC = Negative Control (0.5% DMSO and 0.5% Tween 20) 
; A = 30% oil extract ; B = 60% oil extract; C = 90% oil extract; PC 
= Positive Control (0.5mg/L ciprofloxacin)  
 

                  

Figure 3: Effect of Terminalia catappa seed oil on Aspergillus 
niger: NC = Negative Control (0.5% DMSO and 0.5% Tween 20) ; 
A = 30% oil extract ; B = 60% oil extract ; C = 90% oil extract; PC 
= Positive Control (0.5mg/L ciprofloxacin) 

Discussion 

The seed oil extract of Terminalia catappa was found to have 
tannins, flavonoids and glycosides; but had no inhibitory or 
antimicrobial activity against the micro-organism isolates 
(Escherichia coli, Staphylococcus aureus, and Apergillus 
niger) this was confirmed by its failure to exert any form of 
inhibition zone around the wells containing the oil. This 
could be due to the fact that the concentration of the 
phytochemicals present in the oil was not enough to exert 
antimicrobial activity against the organisms22 and the low 
concentration of the phytochemicals could be due to the 
soxhlet method used in extracting the oil, which has been 
reported to be less effective than methods like steam 
distillation.23 Miksusanti et al (2019)24 reported that 0.5% 
Terminalia catappa seed oil showed no antibacterial effect 
on  Streptococcus mutans (with inhibition zone of 0 mm2), 
but on ethanolysis of the oil the resultant product had 
antimicrobial effect on the same isolate (with inhibition zone 
of 183 mm2).The lack of antimicrobial activity of the oil may 
also be as a result of its inability to diffuse through the 
medium properly due to its poor solubility.22,23  
Other parts of Terminalia catappa have however shown 
antimicrobial activities. Studies by Babayi et al. (2004)25 
using leaf extracts of Terminalia catappa containing 
saponins, tannins and steroids had antimicrobial activity 

against Escherichia coli and staphylococcus aureus which 
are well known to be involve in cutaneous diseases, wounds, 
burns and other different ailments. The antimicrobial activity 
of the leaf extracts of Terminalia catappa may be due to the 
presences of saponins and steroids present which are absent 
in the seed oil. Studies by Pawar and Pal (2002)26 on the 
chloroform extracts of Terminalia catappa root also show 
good antimicrobial activity against Gram-positive and Gram-
negative microorganisms.  
 

Conclusion 

The phytochemical analysis carried out on the oil extract 
revealed the presence of tannins, flavonoids and glycosides 
and the absence of saponins, alkaloids and phenols. 
The antimicrobial investigations carried out on Terminalia 
catappa seed oil base on this study prove had no 
antimicrobial activity against Escherichia coli, Aspergillus 
niger, and Staphylococcus aureus.  
 

Recommendations 

Though Terminalia catappa oil had no significant effect on 
the organisms tested in this research, it may be further tested 
on other microorganisms at different concentrations.  
Adoption of more suitable and effective method of extraction 
and isolation of active compounds such as stem distillation 
and column chromatography is suggested.  
Other uses of the oil should be investigated; such as the 
possibility of its use as a cooking oil and in the manufacture 
of cosmetics. 
 

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