Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 337

Research Article
Phytochemical, antimicrobial and antioxidant studies of Cocos nucifera (L.) flowers
Chinnasamy Chitra Vadivu1*, Mallipalayam Palanisamy Ambika Devi1, Veluchamy Balakrishnan2,
Thirumalaisamy Sundari3
1
PG and Research Department of Botany, Vellalar College for Women (Autonomous), Thindal, Erode- 638012, Tamil Nadu, India
2
PG and Research Department of Botany, Arignar Anna Government Arts College, Namakkal –637 002, Tamil Nadu, India
3
Department of Chemistry, K.S.R. College of Engineering, Tiruchengode-637 215, Tamil Nadu, India

Received: 3 September 2020 Revised: 13 October 2020 Accepted: 30 October 2020

Abstract
Background: Medicinal plant have various phytochemicals and used for the treatment of different kinds of disease.
Objectives: To determine the phytochemical, antimicrobial activity and antioxidant analysis of Cocos nucifera (L.)
flowers. Materials and Methods: Cocous nucifera flowers were shade dried and extracted with solvents ethanol and
water then extract screen for antimicrobial and antioxidant activities. Phytochemical screening such as carbohydrates,
alkaloids, steroids, phlobatannins, saponins, tannins, terpenoids, quinones, flavonoids, phenols and glycosides. For
antimicrobial studies seven bacterial and two fungal clinical isolates selected for the study. DPPH antioxidant studies
also performed based on the phytochemicals present in Cocos nucifera flowers. Results and conclusion:
Phytochemical analysis confirmed that ethanol, chloroform and aqueous extracts were showed the presence of
carobohydrates, flavonoids, phenols, terpenoids, quinones, sterols and glycosides. The Cocous nucifera flower extract
tested seven bacterial clinical isolates and two fungal clinical isolates. The 4mg of plant sample showed the maximum
zone of inhibition. The DPPH assay showed maximum 5 mg of plant extracts having better activity in ethanol,
chloroform and ascorbic acid. The results showed that phytochemical constituents present in Cocous nucifera flower
extract showed that the potential antimicrobial and antioxidant activities.
Keywords: Antioxidants, clinical isolates, DPPH assay, Cocous nucifera, antimicrobial

Introduction potential of medicinal plants as antioxidants in contracting

Free radicals contribute to more than one hundred disorders in such free radicals, induced injury (Pourmorad et al., 2006)
humans including arthritis, ischemia, central nervous system besides well known and conventionally used natural
injury, gastritis, cancer and AIDS (Kumpulainen and Salonen, antioxidants from teas, wines, fruits, vegetables and spices.
1999; Cook and Samman, 1996). Currently available synthetic Antioxidant estimated in Gloriosa superba by several
antioxidants like butylated hydroxyl anisole (BHA), tertiary authors (Gopi, 2016; Jothi et al., 2019; Kavitha and Uduman
butylated hydroquinone and Gallic acid ester, have been suspected Mohideen, 2018). Medicinal plants are considerably useful
to cause or prompt negative health effects. Hence, tough and economically essential. They contain active constituents
restrictions have been placed on their utilization and there is a trend that are used in the treatment of many human diseases (Stary
to alternate them with naturally occurring antioxidants. Moreover, and Hans, 1998).
those synthetic antioxidants also show low solubility and moderate Cocos nucifera (L.) is a significant member of the family
antioxidant activity (Barlow, 1990). Arecaceae (Palm family) popularly known as Coconut,
Recently there has been a boom of importance in the therapeutic coco-da-bahia, or Coconut-of-the-beach. The plant is
basically from Southeast Asia (Malaysia, Indonesia and the
*Address for Corresponding Author: Philippines) and the islands between the Indian and Pacific
Dr. Chinnasamy Chitra Vadivu Oceans (Lima et al., 2015). Every part of it is advantageous
PG and Research Department of Botany, to mankind for numerous functions including food, drinks,
Vellalar College for Women (Autonomous), Thindal, Erode- 638012, fibers, building materials and chemicals discovering their
Tamil Nadu, India way into an immense range of modern day products. The
Email: chitravadivuchinnu@gmail.com coconut maintain a source of meat, milk, oil, fibers,
DOI: https://doi.org/10.31024/ajpp.2020.6.5.4
2455-2674/Copyright © 2020, N.S. Memorial Scientific Research and Education Society. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).

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 Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 338

vitamins and minerals provides enormous health assistance using a sterile cotton swab, to ensure the spread of the tested
beyond its nutritional content. microbes on the surface of the plate completely. Inoculums
8
It is used to be antiblenorrhagic, antibronchitis, febrinfugal and were 10 CFU/ml of bacteria. The 6mm diameter of well was
antigingivitic, antibacterial activity. In ayurvedic medicine, the milk, made with borer on the agar plates. Different concentrations of
oil, cream, water of the coconut is all used to treat hair loss, burns plant extracts were filled in well with the help of micropipette
and heart problems (Manisha and Shyamapada, 2011). The present and one well filled with plant extract. The Ampicillin
work to study about the phytochemical constituents, antimicrobial (10μg/ml) was added in one well and added 100μl of extract in
o
and antioxidant activity of Cocos nucifera flower extract. another well. Incubate the plate at 37 C for 24hrs, after
observed the zone of inhibition. In case of fungal isolates,
Materials and Methods
Itracanazole was added as control and incubated at room
Collection and identification of plant material temperature for 48 hrs.
The flowers of Cocos nucifera was collected from Mallipalayam, Collection of clinical isolates
Gobichettipalayam area, Tamil Nadu, India. These flowers were
In this study, 7 bacterial genera and 2 fungal genera were
shade dried and powered and then stored in air lock covers or
procured from Microtech Microbiology laboratory,
bottles. It is stored for further uses. Medicinally important plant
Coimbatore. All isolates were inoculated into chromogenic
species of Cocos nucifera flowers are selected for this study. The
media for confirmation. According to colony morphology
botanical identification of the plant samples was carried out by
isolates were confirmed as E. coli, K. pneumoniae,
Botanical Survey of India, Coimbatore, India. Certificate No:
E.feacalis, P. aeruginosa, Salmonella sp, Bacillus spp. and
BSI/SRC/5/23/2018/TECH./563.
P. mirabils. in case of fungal isolates, A. niger and Fusarium
Preparation of extracts sp were collected and confirmed with colony morphology.
The collected plant parts were shade dried to remove the water In this study, totally 9 isolates were obtained and subjected
content from the plants to get dried powder. The dried plant for this investigation.
flowers were extracted with solvents like ethanol and water for Antioxidant activity - DPPH Assay
antimicrobial and antioxidants activities. The powered extract
Aliquot 3.7 ml of absolute methanol in all test tubes along
was extracted by taking 5g of sample in 100ml of solvent. The
with blank was taken. Then, added 100µl of absolute
mixture was kept in shaking condition for about 24 to 48 hours
methanol to blank.100 µl of Ascorbic acid to tube marked as
by closing it tightly. This is because some of the solvent gets
standard and 100 µl of respective samples to all other tubes
evaporated quickly. Then they were taken and filtered using
marked as tests. Finally, added 200µl of DPPH reagent to all
Whatmann No.1 filter paper. These filtered extracts were dried
by pouring it in petridishes and allow them for dry up to one the test tubes including blank. Then, all test tubes were
week. The dried plates were than scarped completely using incubated at room temperature and dark condition for
sterile blades. The collected powder was taken and stored in minimum of 30 minutes of duration. The absorbance of all
proper containers and then sealed using parafilm. samples was taken in Spectrophotometer at 517nm (Naznin
and Hasan, 2009).
Test pathogens
Results
The clinical isolates of E. coli (Mm306), P. aeruginosa
(MM502), S. aureus (MM1002), Salmonella sp (MM104), The results of phytochemicals screening of Cocos nucifera
Bcillus sp (MM919), E. feacalis (MM611), K. pneumoniae flowers showed the presence of various phytochemicals
(MM218), A. niger (MM316), Fusarium sp (MM308) were (Table 1). Three solvents such as ethanol, chloroform and
procured from Microtech, Microbiology Laboratory, Aqueous showed positive results for the presence of
Coimbatore which were used for the present study. All isolates flavonoids, phenols, tannins and carbohydrate, sterols and
were inoculated onto Chromogenic agar media for terpinoids. In addition, the ethanol extract showed the
confirmation. After 24 hrs, observed pigment and colony presence of saponins and quinones. The alkaloids observed
morphology of each isolates. According to color and colony only in chloroform extract. The proteins are absent in all of
morphology, isolates were confirmed. the extracts and ethanol extract had highest number of
phytochemicals and followed by chloroform extract.
Determination antimicrobial activity of plant extract
Collection of clinical isolates
This test was carried out according to the method of Jahir et al.
(2011). The plates were inoculated with freshly prepared over According to colony morphology isolates were confirmed
night inoculums which were swabbed over the entire surface of the as E. coli, K. pneumoniae, E. feacalis, P. aeruginosa,
medium, rotating the plate 60 degrees after each application by Salmonella sp, Bacillus spp. and P. mirabils. in case of

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 Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 339

Table 1. Preliminary phytochemicals analysis of Cocos nucifera fungal isolates, A. niger and Fusarium sp were collected and
flowers confirmed with colony morphology. The results were
S. No. Phytochemicals test Chloroform Extract Ethanol Extract Water extract tabulated in table 2 and figure 1. In this study, totally 9
1. Alkaloids + - - isolates were obtained and subjected for this investigation.
2. Carbohydrate + + +
3. Flavonoids + + +
Antibacterial and antifungal activities of ethanol extract
4. Phenols + + + of Cocos nucifera L.
5. Saponins - + -
6. Tannin + - + According to results of phytochemicals analysis, highest
7. Terpenoids + + + phytocompounds containing ethanol and chloroform
8. Quinones + + +
extracts were selected for the antimicrobial activity. In the
9. Sterols + + +
10. Proteins - - - present study, 7 bacterial genera were subjected to
11. Glycosides + + + antimicrobial activity test with both solvents extracts. In
+, Present; - Absent
case of ethanol extract, highly suppressed to S. aureus and
followed by Bacillus spp, E. coli. Among the 7 genera,
Table 2. Morphological characterization of bacterial isolates
Salmonella was highly resistance to ethanol extract. The
S. No. Name of the Microorganisms Morphology/Colour
zone of inhibition was ranged from 10mm to 22mm. In this
1. E. coli Pink colony chromogenic media study, most of the isolates were suppressed while using 3mg
2. Pseudomonas aeruginosa Colour less colony chromogenic media concentration of extract (Table 3 and Figure 2). In case of
3. Enterococcus faecalis Small blue colony chromogenic media
fungal genera, A. niger was highly suppressed, it exhibiting
4. Klebsiella pneumonia Blue colony with mucoid chromogenic
zone of inhibition was 14 mm to 19 mm, and zone of
media
inhibition was stated with 1mg concentration of extract. The
5. Bacillus milky, large, convex, opaque, dry colonies
Fusarium sp. was not suppressed with ethanol extract
6. Salmonella sp. Black colour on SS agar
7. S. aureus Yellow colour on chromogenic media (Table 4; Figure 3).
8. A. niger Black colony on SDA media Antibacterial and antifungal activities of chloroform
9. Fusarium sp. Pale colored; whitish to yellow extract of Cocos nucifera L.
In this study, chloroform extract of Cocos nucifera also
utilized for the antimicrobial activity. The zone of inhibition

Table 3. Antibacterial activity of ethanol extract of Cocos

nucifera Linn.
S. Name of the Different Concentration of plant extract
No. Microorganisms (mg) (Zone of inhibition in mm)

1 2 3 4 5 6

1. E. coli 14 19 17 19 - -
2. K. pneumoniae - - 14 17 - -
3. P. aeruginosa - - 14 15 - -
4. Salmonella sp. - - - - - -
5. S. aureus 10 14 19 22 - -
6. E. faecalis - 10 15 17 - -
7. Bacillus sp. 15 17 19 21 - -

Table 4. Antifungal activity of ethanol extract of Cocos

nucifera L.

S. Name of the Different Concentration of plant extract

No. Microorganisms (mg) (Zone of inhibition in mm)

1 2 3 4 5 6

1. A. niger 10 12 14 16 - -
2. Fusarium sp. - - - - - -
Figure 1. Confirmation of bacterial and fungal isolates

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 Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 340

Figure 3. Antifungal activity of ethanol extract of Cocos

nucifera L.

Figure 2. Antibacterial activity of ethanol extract of Cocos

nucifera L.

Table 5. Antibacterial activity of chloroform extract of Cocos

nucifera L.

S. Name of the Different Concentration of plant

No. Microorganisms extract (mg) (Zone of inhibition in
mm)

1 2 3 4 5 6

1. E. coli 12 13 15 17 - -
2. K. pneumoniae - - 12 14 - -
3. P. aeruginosa - - - 12 - -
4. Salmonella sp. - - - 12 - -
5. S. aureus - 10 11 14 - -
6. E. faecalis - 10 13 14 - -
7. Bacillus sp. - 10 11 14 - -

was ranged from 10mm to 17mm. among the 7 bacterial genera;

E. coli was highly suppressed and followed by E. faecalis. The
Figure 4. Antibacterial activity of chloroform extract of
Salmonella spp. was not suppressed with chloroform extract. In
Cocos nucifera L.
this study, 5 isolates were suppressed while using 3mg of
concentration and all isolates were suppressed while using 4mg
concentration of plant extract (Table 5 and Figure 4). In case of Antioxidant activity of ethanol extract of Cocos nucifera
fungal genera, both isolates were minimal suppressed with Linn.
chloroform extract (Table 6 and Figure 5). In this investigation, According to phytochemicals constituents, ethanol and
relevant solvents and standard antibiotic of ampicillin were used chloroform extracts was utilized for the antioxidant activity
for control. These were not produced the zone of inhibition through DPPH free radical scavenging assay. The
against bacterial and fungal genera. antioxidant capacities of the extracts were compared with

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 Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 341

Table 6. Antifungal activity of chloroform extract of Cocos

nucifera L.

S. Name of the Different Concentration of plant extract (mg)

No. Microorganisms (Zone of inhibition in mm)

1 2 3 4 5 6

1. A. niger - - - 10 - -
2. Fusarium sp. - - - - 11 -

Figure 6. Percentage of antioxidant activity of Cocos

nucifera L. extract

be 3mg/ml, 3.6mg and 2mg/ml respectively (Table 7).

Discussion
In this study, ethanol and chloroform extracts were used for
the antimicrobial activity. Among them, ethanol showed
better activity then chloroform extract. In case of ethanol
extract, highly suppressed to S. aureus and followed by
Bacillus sp, E. coli. Among the 7 genera, Salmonella was
highly resistance to ethanol extract. Similarly Cristiane et al.
(2016) were determined the antibacterial activity of flowers
of Cocos nucifera against S. aureus and P. aeruginosa. In case
of chloroform extract, E. coli was highly suppressed and
followed by E. faecalis. The antimicrobial activity of ethanol
extract of Cocos nucifera against bacterial isolates. In Allium
Figure 5. Antifungal activity of chloroform extract of Cocos
species antimicrobial studies carried out (Semerci et al.,
nucifera L.: (1) 1mg; (2) 2mg; (3) 3mg; (4) 4mg; (5)
2020).
Chloroform; (6) Itracanazole (5mcg)
Several investigations on medicinal plants indicate that
ascorbic acid as standard antioxidant. The graphical organic solvents such as methanol are extensively used for
representation shows the increase in DPPH activity with crude extraction before being re-extracted to obtain purified
response to the increase in drug concentration (Fig 6). Ethanol active compounds using some other organic solvents. Present
extract of Cocos nucifera exhibited a greater antioxidant effect study also agreed with previous studies, which was inhibited
at 5mg/ml (78.66%) compared with the effect of ascorbic acid at by most of the isolates. The present study was first
the same concentration (89.48%). In case of chloroform extract, investigation, in our literature knowledge, this is the first
highest scavenging rate was occurred on 5mg with 71.58%. The study, and no one inhibited the biofilm and betalactamase and
IC50 value of ethanol, chloroform and ascorbic acid was found to MDR isolates with Cocos nucifera.

Table 7. Antioxidant activity of Cocos nucifera L.

S. No. Concentration (mg/ml) DPPH free radical scavenging effect (%)

Ethanol extract Chloroform extract Ascorbic acid

1. 1 15.62 10.20 29.23

2. 2 25.14 22.13 49.79
3. 3 49.50 36.44 64.35
4. 4 68.12 60.42 79.91
5. 5 78.66 71.58 89.48

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 Asian Journal of Pharmacy and Pharmacology 2020; 6(5): 337-343 342

It has observed that the natural compounds in higher plants have Conclusion
antioxidant activity. Several mechanisms have been proposed to The antimicrobial activity study revealed that ethanol
be involved in the antioxidant activity such as hydrogen extract was highly suppressed to the S. aureus, Bacillus and
donation, termination of free radical mediated chain reaction, E. coli but Salmonella sp. was resistant. In case of fungi,
prevention of hydrogen abstraction, chelation of catalytic ions, minimal inhibition zone was observed from A. niger and
and elimination of peroxides (Gordon, 1990). The unsettle to the Fusarium .In case of Chloroform extract, this was highly
complex reactive nature of phytochemicals, the antioxidant effective against E. coli followed by E. faecalis. The
activities of plant extracts can be evaluated by DPPH method, Salmonela sp. and the fungal isolates of A. niger and
authenticity (Schlesier et al., 2002; Chanda and Dave, 2009). Fusarium were lesser zone inhibition than E. coli and E.
Therefore in the present study, the antioxidant activities of Cocos faecalis. Ethanol extract of Cocos nucifera exhibited a
nucifera extract were analyzed by DPPH free radical scavenging greater antioxidant effect and highest scavenging rate was
activity. Free radicals are involved in many disorders like occurred in chloroform extract.
neurodegenerative diseases, cancer and AIDS. Antioxidants
Acknowledgement
have scavenging activity and it's significantly contributed for the
treatment of diseases. DPPH stable free radical method and The authors are thankful to the Management, Principal and
which is useful for the determination of antioxidants in plants Head of the Department of Botany, Vellalar College for
(Koleva et al., 2002; Suresh et al., 2010). Women (Autonomous) Thindal, Erode for providing
necessary laboratory facilities to carry out the work
In this study, two solvents of plant extracts were utilized for the
successfully.
antioxidant activity, among them, ethanol showed better activity
than other extracts. The IC50 value of ethanol, chloroform and Conflicts of interest
ascorbic acid was found to be 3mg/ml, 3.6mg and 2mg/ml We declare that we have no conflicts of interests
respectively. This activity result was resembled to ascorbic acid
Ethical clearance
activity. Similarly, Cocos nucifera contain antioxidants having a
different chemical nature from ç-Sitosterol, Stigmast, quercetin Not necessary for this work.
content was found to be high in Cocos nucifera flowers, and Source of funding
capable to confer a high antioxidant power to this species (Singla This work did not received and fund from any funding
et al., 2011). Since DPPH assay is easily reproducible and agency
linearly related to molar concentration of the antioxidants
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