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Kongunadu Research Journal 10 (1): 48-52, 2023 ISSN 2349-2694, All Rights Reserved,
Publisher: Kongunadu Arts and Science College, Coimbatore. https://krjournal.com/index.php/krj/article/view/609

RESEARCH ARTICLE
Phytochemical, Antioxidant potential and ftir analysis on the matured leaves of Camellia
Oleifera abel

Vasini, V.
a
, Betty, T.
a
, Malini, R.P.
a
and Sumathi, P.
a
*
a
Department of Botany, Kongunadu Arts and Science College (Autonomous), Coimbatore - 641029,
Tamil Nadu, India.

ABSTRACT
Phytochemicals are physiologically active compounds and are derived from plants. Majority of the
phytochemicals have been known to bear therapeutic activities like antibacterial, antioxidant, antifungal,
antispasmodic, anticancer, hepatoprotective etc. The aim of this study is to evaluate the phytochemicals in the
matured leaves of Camellia oleifera belonging to the family Theaceae. The phytochemicals were extracted
from the dried matured leaves of C. oleifera using solvent extraction method. The preliminary phytochemical
analysis showed the presence of alkaloids, tannins, saponins, phenols and flavonoids in the ethanolic extract.
Further the antioxidant property of the leaf extract was carried out using standard procedure. The extract
was subjected to FTIR analysis. The study shows that the ethanolic extracts possess secondary metabolites
and antioxidant properties that are therapeutically valuable.
Keywords: Camellia oleifera, secondary metabolites, antioxidant, FTIR.
1. INTRODUCTION
Plant extracts have great potency and can be
used for a variety of purposes. Approximately 80%
of the world’s population relies on traditional
medicine for health care, and most therapies use
plant extracts and their active compounds [1].
Medicinal plants are the richest bio-resources of
folk medicines and traditional systems of medicine,
and food supplements, pharmaceuticals industries
and chemical entities for synthetic drugs [2].
India is the birth place of renewed system
of indigenous medicine such as Siddha,
Ayurvedha and Unani. Plants are a good source of
various functionally active secondary metabolites
and a good source of essential nutrients [3]. Recent
studies have demonstrated the nutritional and
nutraceutical efficacy of different plant tissues
proving their commercial value [4, 5]. Several
techniques are available to identify the
phytochemical compounds in plant extracts. For
example, Fourier transform infrared spectroscopy
(FTIR) is a method used to identify the functional
groups in gaseous, liquid and solid materials via
infrared radiation beams [6]. It is possible to detect
the minor changes in the primary and secondary
metabolites in leaves by observing the IR spectra
[7]. FTIR has been used to identify the complicated
structures of plant secondary metabolites and in
the characterization of bacterial, fungal and plant
species [8]. This technique presents a rapid,
inexpensive, and rather non-invasive method for
obtaining chemical characteristics of a biological
sample.

2. PLANT DESCRIPTION:
Kingdom: Plantae
Phylum : Tracheophyta
Order : Ericales
Family : Theaceae
Genus : Camellia L.
Species : Camellia oleifera Abel.

General Description:
Evergreen. Shrub reaching upto 5m in
height. Leaves 3–7 cm in length, 1.2–3cm in width,
broad, elliptic with a finely serrated margin. The
flowers are dark pink, 5–8 petals and 5–7 cm in
diameter.


*Correspondence: Sumathi, P. Department of Botany, Kongunadu Arts and Science College, Coimbatore - 641029, Tamil Nadu, India.
E.mail: [email protected]

DOI:10.26524/krj.2023
.
7

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Figure 1. Habit of Camellia oleifera Abel.

3. MATERIALS AND METHODS

3.1. Collection of the plant samples
The matured leaves of Camellia oleifera
belonging to the family Theaceae was collected
from Kovillatty Village, Manjoor, Nilgiri district,
Tamil Nadu, India during the month of June 2022,
identified and confirmed by the Flora of the
Presidency of Madras [9]. The collected materials
were washed thoroughly with tap water to remove
the sediment particles. The plant leaves were
shade dried and powdered coarsely. The powder
was stored in air tight container and used for
further successive extraction.

3.2. Solvent extraction
The powdered plant sample was extracted
with sequential solvent analysis (viz, Hexane,
diethyl ether, ethanol and aqueous solutions) using
cold maceration method for 8 to 10 hours in order
to extract the non-polar and polar compounds. The
solvent of respective extract was reduced at room
temperature and stored under 4⁰C for further use.

3.3. Phytochemical screening
Chemical tests were carried out using
standard procedure to identify the preliminary
phytochemical screening and the ethanolic extract
were analyzed quantitatively. [10-15]

3.4. Determination of in vitro antioxidant activity

3.4.1. DPPH radical scavenging activity
The hydrogen donating capacity was
assessed by using stable DPPH method. Briefly, a
solution of 0.1Mm DPPH was prepared using
methanol. The sample (50-250μg/mL) was mixed
with 5.0 mL of DPPH solution. Reaction mixture
was shaken incubated at 30⁰ c for 20 minutes and
the absorbance was measured at 517nm. Results
were compared with the activity of rutin.
Antioxidant activity of the extract were expressed
as IC50, the values were calculated from the linear
regression of the percentage antioxidant activity
versus concentration of the extracts. A lower IC50,
values indicate greater antioxidant activity. DPPH
dis-colouration of the sample was calculated using
the formula.
DPPH radical scavenging activity (%) = [(control
OD- sample OD)/control OD] x100.

3.4.2. ABTS
+
free radical scavenging assay
Antioxidant activity was performed using an
improved ABTS+ method proposed by Siddhuraju
and Manian. The ABTS radical cation was ABTS+
was generated by a reaction of 7mm ABTS+ and
2.45 mm potassium persulphate and the mixture
was incubated for 12 – 16 hrs at room temperature
in dark. Prior to assay, the solution was diluted in
ethanol (about 1:89 v/v) and equilibrate to obtain
an absorbance of 0.700± 0.02 at 734 nm. 10 μL/ml
of the sample was added to 1.0ml of diluted ABTS+
solution. After 30 min of incubation, absorbance
was read at 734nm. Trolox was used as reference
material.

3.5. FTIR analysis: Fourier Transform Infra-Red
Spectroscopy Analysis
The FTIR spectra of the fractions with the
highest anti-inflammatory activity were carried out
using FTIR-8400S spectrophotometer (Shidmazu
model). FTIR analysis of the ethanolic extract was
performed using Perkin Elmer Spectrophotometer
system, which was used to detect the characteristic
peaks and their functional groups. The peak values
of the FTIR were recorded. Each and every analysis
was repeated twice and confirmed the spectrum.
Fourier Transform Infra-Red (FTIR) is a tool used
to identify the type of chemical compound in the
sample. The sample was loaded in the FTIR with
the scan range from 400-4000cm-1 with a
resolution of 4cm-1 and the results were recorded.

4. RESULTS AND DISCUSSION

4.1. Qualitative estimation of secondary metabolites
Secondary metabolites are chemicals
produced by plants that allow them to compete in
their natural environment. In present investigation
secondary metabolites were analyzed both
qualitative and quantitatively. The present analysis
of phytochemical screening revealed that C. oleifera
contained flavonoids, phenols and tannins where

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present. Whereas saponin and steroids were
absent in the ethanolic extract.
4.2. Quantitative estimation of secondary
metabolities
Flavonoids are a large class of natural
aromatic compounds as there is reported to be the
most common plants’ phenolics [16]. The biological
and oxidative properties of flavonoids are
responsible for their anti-allergic, cardioprotective,
anti-diabetic, anti-inflammatory, anti-oxidative
activity, and free radical scavenging capacity [17].
The total flavonoid content of the extract was
found to be 1.143±0.04mg RE /g in ethanolic
extract and was expressed in Rutin equivalent.
Polyphenols are secondary plant
metabolites that play a vital role in protecting
plants from UV radiation and disease attacks [18].
The phenolic content in the extract of C. oleifera
were expressed in gallic acid equivalence and were
found to be 0.933± 0.03 mg GAE /g in ethanol
.They carry strong natural antioxidants having key
role in wide range of biological and
pharmacological properties such as anti -
inflammatory, anticancer, antimicrobial, anti-
allergic, antiviral, antithrombotic, hepatoprotective
[19] .Tannins are the main
polyphenolics distributed widely in the range of 5
to 10% of dry vascular materials found mainly in
bark, stems, seeds, roots, buds, and leaves [20]. The
total tannin content of the extract was expressed in
Tannic acid equivalents and was found to be
1.111±0.01mg GAE/g in ethanolic extract. In the
past few years, tannins have also been studied for
their potential effects against cancer through
different mechanisms [21].
For FTIR, the analysis time was less than
five minutes and it required a minute quantity of
the sample [22]. The FTIR spectrum was used to
identify the functional group of the active
components based on the peak value in the region
of infra-red radiation. The functional groups of the
components were separated based on its peak
ratio. As illustrated in (Figure 1), FTIR spectrum of
ethanolic extract showed different peaks at 617.55,
628.99, 1057.85, 1383.78, 2344.42, 2927.66,
3305.05, 3362.24cm-1. It was confirmed the
presence of functional groups such as Halo
compound (C-Br streching), Halo compound (C-Br
streching), sulfoxide (S=O streching), Phenol (O-H
binding), Carbon dioxide (O=C=O), Aldehyde (C-H
streching), Alkyne (C-H streching) and Aliphatic
primary amine (N-H streching).

Table 1. Qualitative phytochemical analysis of
ethanolic matured Leaf extract of C. oleifera

Constituents Presence/
absence
Alkaloids ++
Flavonoids +++
Phenols +
Tannins +
Saponins -
Steroids -

Table 2. Quantitative analysis of ethanolic
Matured Leaf extract of C. oleifera

Constituents Ethanolic extract
mg/g ±SD
Flavonoids 1.668±0.02
Phenols 0.933±0.03
Tannins 1.111±0.01


Table 3. Antioxidant activity of ethanolic leaf
extract of C. oleifera

ASSAY IC50 value
DPPH 0.118±0.02
ABTS 0.156±0.03


Figure 2. FTIR Spectrum of ethanolic Matured
leaf extract of C. oleifera

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4. CONCLUSION
The result of this study shows the
presence of some phytochemicals such as alkaloids,
flavonoids, phenols and tannins in ethanolic leaf
extracts of C. oleifera. The flavonoids, phenols and
tannins were analyzed quantitatively. The
antioxidant activity revealed the highest inhibitory
concentration in the ethanolic extract. The FTIR
analysis confirmed the presence of various
bioactive compounds. Thus it is concluded that the
plant has various medicinal properties and can be
used for further pharmacological studies.

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