Advances in Science Engineering and Technology
, ISSN: 2321-9009
Volume- 3, Issue-4, Oct.-2015
Qualitative And Quantitative Analysis Of Medicinal Tree’s Belonging To Myrtaceae Family
S.B.NasrinFathima, Department of plant biology and biotechnology,
The Principal, GovernmentAarts and Science College, Kovilpatti,
species of Myrtaceae family such as Syzygium densiflorum, Syzygium tamilnadensis and Euginea candolleana,
which Syzygium densiflorum is categorized as a vulnerable species in the IUCN red list of threatened species.
Preliminary phytochemical analysis revealed the presences of tannins, alkaloids, and phenols, whereas saponins,
of total phenol, total flavonoids and Tannin content in n-hexane, ethyl acetate and methanol extracts of Syzygium
methanolic extract shows high phenolic, flavonoid and tannins content for Euginea candolleana. The Syzygium
high in Syzygium tamilnadensis of n- hexane and ethyl acetate extract. This report shows the complete study on
qualitative and quantitative analysis of the tree leaves extracts.
Keywords— Syzygium Tamilnadensis, Syzygium Densiflorum, Euginea Candolleana.
Syzygium densiflorum ,Syzygium tamilnadensis and
Euginea candolleana belong to the family Myrtaceae.
The Myrtaceae is the myrtle family, of dicotyledonous
plants placed within the order myetales. Myrtle, clove,
guava and eucalyptus belong to this family.
are woody, with
. Recent estimates
includes over 5650 species
occurring in some 130-150 genera, .
Plants of the
Family Myrtaceae are predominantly from the
Southern Hemisphere and are found in tropical,
and temperate Australia.
densiflorum has been categorized as a vulnerable
under IUCN red list of threatened species and is
endemic to India (IUCN 1998). There are over 150
Genera in the family and approximately 50 per cent of
these are Australian. Plants from the Myrtle Family
share common characteristics; including aromatic
leaves with oil glands and may be either shrubs or
trees. Syzygium tamilnadensis is a tree up to 15m tall
and is endemic to Western Ghats. Its leaves are simple,
opposite and decussate. Its flowers are small in
terminal corymbose cyme .
It is a tree, native from Atlantic rain
names cambuí roxo
It is quite rare in the wild,
and has seen limited use in landscaping for its bright
The fruits are
tree have been used for the treatment of pain and fever
. They have oil bearing leaves which contain
isomers of guaiol and cadinol, δ-elemene and
Medicinal plants contain some organic compounds
human body and these bioactive substances include
tannins, alkaloids, carbohydrates, terpenoids, steroids
and flavonoids . These compounds are synthesized
by primary or rather secondary metabolism of living
organisms. Secondary metabolites are chemically and
taxonomically extremely diverse compounds with
obscure function. They are widely used in therapy,
veterinary, agriculture, scientific research and other
countless areas .
Phytochemical is biologically active, naturally
plants such as barks, leaves, flowers, roots, fruits, and
seeds . In the present work, qualitative and
quantitative analysis was carried out in three
leaves of tree species, (Syzygium
ethyl acetate and methanol) for the better yield of
biologically active compounds.
Fresh leaves of Syzygium tamilnadensis, Syzygium
Wayanad district, Kerala. They were identified and
authenticated at “Gandhigram Rural Institute,
Dindigul, Tamilnadu, by Dr. Ramasubbu, Department
Of Plant Biology and Plant Biotechnology”. The plant
materials were shade dried for the water molecules to
get evaporated. Once dried, they were coarsely
powdered and transferred to air tight container for
The Crude leaf extract was prepared by percolation
extraction method. About 100 gms of powered extracts
were extracted three times by cold percolation method
with 300 ml of n-hexane, ethyl acetate and methanol at
room temperature for 72 hours. The filters were
concentrated under reduced pressure at 40°C and
stored in refrigerator at 2-8°C for use in subsequent
The extracts were screened for the presence of
Carbohydrates, Tannins, Saponins, Flavonoids,
Triterpenoids, Phenols, Coumarins , Phytosteroids,
Phlobatannis, and Anthraquinones for all the three
leaves samples of (Syzygium densiflorum, Syzygium
solvents (n-hexane, ethyl acetate and methanol)
Total phenol content in n-hexane, ethyl acetate and
methanol extracts of all the three leaves samples was
determined by Folin-Ciocalteau method  with
some modifications. Briefly, 0.1ml of extract (200, 600
and 1000 µg/ml), 1.9ml distilled water and 1 ml of
Folin-Ciocalteau’s reagent were seeded in a tube. Then
1 ml of 100g/l Na
mixture was incubated at 25° C for 2 hours and the
absorbance of the mixture was read at 765 nm. The
sample was tested in triplicate and a calibration curve
with six data points for Catechol was obtained. The
results were compared to a Catechol calibration curve
and the phenolic content of all the three leaves samples
was expressed as mg of Catechol equivalents per gram
Total Flavonoid content:
Total flavonoid content in n-hexane, ethyl acetate and
methanol extracts of all the three leaf samples was
determined by calorimetric method as described in the
literature. 0.5ml of this sample was mixed with 2 ml of
distilled water and subsequently with 0.15ml of
solution (15%). After 6 min, 0.15 ml of AlCl
solution (10%) was added to the mixture. Immediately,
water was added to bring the final volume to 5 ml and
the mixture was thoroughly mixed and allowed to
stand for another 15 min. Absorbance of the mixture
was then determined at 510 nm versus prepared water
blank. Results were expressed as quercetin equivalents
(mg quercetin/g dried extract) .
Total Tannin content in n-hexane, ethyl acetate and
determined by Sun et al., 1998. To 50µl of properly
diluted sample, 3 ml of 4% methanol vanillin solution
and 1.5 ml of concentrated hydrochloric acid were
added. The mixture was allowed to stand for 15 min
and the absorption was measured at 500 nm against
methanol as a blank. The amount of total condensed
tannins is expressed as mg (+) - catechin g
. All the
5gms of the sample were weighed into a 250 ml beaker
and 200 ml of 10% acetic acid in ethanol was added
and covered and allowed to stand for 4 hours. This was
filtered and the extract was concentrated on a water
bath to one quarter of the original volume.
Concentrated ammonium hydroxide was added drop
wise to the extract until the precipitation was
completed. The whole precipitated solution was
collected and washed with dilute ammonium
hydroxide and then filtered. The residues in the
alkaloids were dried and weighed .
All the experiments were carried out in triplicate and
the results were represented in mean ± standard
The preliminary phytochemical characteristics of
leaves of three medicinal tree species (Syzygium
densiflorum, Syzygium tamilnadensis, and Euginea
candolleana) were tested against three solvents
(n-hexane, ethyl acetate, and methanol) and were
summarized in the [Table-1]. The results revealed the
presence and absence of medicinally active compounds
in the three leaves samples. Tannins, alkaloids and
phenols were present in all the tested extracts of all the
three leaves samples. Saponnins, terpenoids, steroids,
pytosteriods, phlobatannina and anthraquinonol were
absent for the tested extracts.
The presence of such compounds in huge varieties of
be used in the treatment of various diseases .
Table-1. Phytochemical analysis of plant leaves
: Euginea candolleana; (++: Strongly
present, +: Weakly - : Absent).
The amount of total phenols, flavonoids, and tannins
spectrometrically and was summarized in [Table-2, 3
and 4]. The maximum phenol content in the
methanolic extracts of E. candolleana was 463.9mg/g
and of S.tamilnadensis was 335.9mg/g, and of
S.densiflorum was found to be 389.0mg/g. In n-hexane
and ethyl acetate, the maximum phenol content
present in S.tamilnadensis and E. Candolleana was
117.5mg/g and 259.4mg/g respectively. The total
flavonoid content for the methanol extract of
S.densiflorum was 152.8mg/g, and of E. candolleana
was 215.8mg/g but it was absent in S.tamilnadensis.
Ethyl acetate and n-hexane didn’t show flavonoid
content for the leaves samples.The concentration of
flavonoids in leaves extracts depends on the polarity of
solvents used in the extract preparation .The
amount of total condensed tannins in the methanol
extract of S.densiflorum was 589.3mg/g and in E.
acetate extracts of S. tamilnadensis, was 162.1mg/gm,
samples showed the presence of condensed tannins in
ethyl acetate extract, whereas n-hexane does not show
tannin content for S.densiflorum and E. candolleana
and were present in S.tamilnadensis (72.6mg/gm).
Natural antioxidants such as phenols, flavonoids and
tannins are natural disease preventing, health
promoting and anti-ageing substances .
Table-2. Total phenolic content of leaves extracts in
The results represented as mean ± SD of three
The results represented as mean ± SD of three
Table-4: Total condensed tannins of leaves extracts in
It is well-known that phenolic compounds contribute
to quality and nutritional value in terms of modifying
color, taste, aroma, and flavor and also in providing
health beneficial effects. They also serve in plant
defense mechanisms to counteract reactive oxygen
species (ROS) in order to survive and prevent
molecular damage by micro organisms, insects and
herbivores. Condensed tannins can be found
commonly in used foods. They are polyphenolic
compound with bitter taste. Tannins are used in folk
medicine to combat diarrhoea, hemorrhoids and to
heal wounds as bactericides and other poison and
Chloroform, benzene and methanol were used as a
ethyl acetate and methanol as solvent source for the
extraction of metabolites. Since the polarity of
methanol is higher, most of the secondary metabolites
of S.tamildanensis, S. densiflorum and E. candolleana
qualitative tests screened for the presence of secondary
metabolites, four showed positive results for all the
three solvents used. Carbohydrates were present in
showed positive results for S. densiflorum and
extract of E.candolleana. Cardiac glycosides were
present in all the extracts and were absent for S.
densiflorum and E.Candolleana of methanolic
extracts. Quinines and Coumarins tests showed
positive results for methanol extracts.
Preliminary qualitative tests for two algae Gracilaria
four solvents (n-hexane, petroleum ether, ethyl acetate
and ethanol).Sixteen phytochemical tests were done,
in which carbohydrates, quinines, and glycosides
showed positive results for all the solvent extracts and
were absent for triterpenoids, coumarines, proteins,
for all the four solvent extracts. The Eugenia uniflora
was tested against ethanol, methanol, chloroform,
petroleum ether, ethyl acetate, acetone, aqueous cold
and hot. Aqueous hot, ethanol and methanol extract
showed best results for the entire test done . These
compounds present in the variety of medicinal plants
have significant application against human pathogens,
including those that cause enteric infections and are
reported to have curative properties against several
pathogens and therefore could suggest their use in the
treatment of various diseases .
In our study where we used n-hexane, ethyl acetate
and, methanol as solvent extractions, methanol
showed maximum phenol content for all the three
leaves samples of S.tamilnadensis, S. densiflorum and
showed total phenol content for S.tamilnadensis and
but absent for n-hexane and ethyl acetate extract. Ethyl
acetate showed total condensed tannins for all the three
leaves samples. Methanol extraction showed the total
methanol extraction showed best results and this is
because methanol is a polar solvent and it can be easily
The total phenol contents in the examined extracts
ranged from 463.9 to 10.3mg of Catechol equivalents
per gram of extract. The highest concentration of
phenols was measured in methanol and ethyl acetate
extracts, whereas n-hexane extracts contained
considerably smaller concentration of phenol. In the
previous study, the total phenolic content was
examined with methanol, acetone, ethyl acetate and
petroleum ether in Marrubium peregrinum. . The
total phenolic content ranged from 27.26 to 89.78
mg/g of dry weight of extract, expressed as gallic acid
equivalents. The total flavonoid concentrations varied
from 18.72 to 54.77 mg/g, expressed as rutin
equivalents. Methanolic extracts of M. peregrinum
phenolic and flavonoid
concentration.The total phenolic content in Paederia
foetida ranges from 35.5mg/g and Syzygium aqueum
ranges from 20.77mg/gm. The maximum phenolic
content was shown in methanol leaves extracts of both
the plant materials. .
The solvents used for the phenol and flavonoid content
in seven medicinal plants such as Bryophyllum
methanol, ethanol, and acetone . Among all the
seven plants, the total phenol content was found to be
in Tinospora cordifolia (leaves) and the flavonoids
were found to be in Terminalia bellerica (leaves) for
all the solvents used. The total phenol content was
identified, for few Australian herbs such as
Tasmannia pepper berry leaf, Anise myrtle, Lemon
myrtle, Bush tomato, and Wattle seed and methanolic
extract of Tasmania pepper leaf showed maximum
phenolic content .
Phenol and flavonoid content in three plants Primula
were extracted with methanol, distilled water, and
methanol and water mixture. The total phenol and
flavonoid for methanolic extracts of Primula
which it was found that methanol showed maximum
phenol and flavonoid content for all the three leaves
samples. Whereas in Indigofera aspalathoides, the
highest concentration of phenol content and tannins
were obtained in methanol extracts, in which the
phenolic content was 47.38mg/g and the tannin
content was 34.59mg/g .
The quantitative phytochemical analysis of roots of S.
caryophyllatum and S. densiflorum were treated with
three different solvents such as acetone, methanol, and
aqueous. A higher concentration of total phenol
content was noted in the acetone root extract followed
by aqueous and methanol extract of S. caryophyllatum
and very low total phenol content was seen in the
aqueous extract of S. densiflorum. Very low
concentrations of flavonoids were noted in aqueous
root extracts of both the plants and higher
concentration of total flavonoid was noted in the
acetone root extract of S. caryophyllatum. The tannin
content was found to be equal in the acetone, aqueous
and methanol extracts of root. Whereas, the lowest
amount of tannin was noted in S. densiflorum extracts
variation depending upon the extraction and solvent
Tannins are known to possess general antimicrobial
and antioxidant activities . Recent reports show
that tannins may have potential value as cytotoxic and
antineoplastic agents . Phytochemical such as
saponins, terpenoids, flavonoids, tannins, steroids and
alkaloids are reported to have anti-inflammatory
effects -. Phenolic phytochemicals have
anti-inflammtory . Tannins, that contain tannic
molecular weight and are usually divided into two
proanthocyanidins they are found in abundance in
fruits and fruit products. This unique group of phenol
is associated with the astringency and color of fruits
. Moreover, proanthocyanidins are known to
possess some biological properties that are of vital
importance for human health. In the previous study,
fruits from the Ida mountains were collected and
determined for the total condensed tannins, for Sour
cherry the tannin content was 163.4mg/g, for cherry
was 113.1mg/g for apricot was 38.8mg/g and for
greengage plum was 20.42mg/g. The maximum
condensed tannins were found to be in sour cherry
epidermis of the leaf tissues, most common in the
upper epidermis. However, in evergreen plants,
tannins are evenly distributed in all leaf tissues. They
serve to reduce palatability and, thus, protect against
predators. Condensed tannins, also known as
proanthocyanidins, are polymers of 2 to 50 (or more)
flavonoid units that are joined by carbon-carbon bonds,
which are not susceptible to being cleaved by
hydrolysis. Although hydrolyzable tannins and most
condensed tannins are water soluble, few very large
condensed types of tannin are insoluble .Tannins
play a major role as antihaemorrhagic agent and show
to have immense significance as antihypercholesterol,
hypotensive and cardiac depressant properties .
Previously, researchers took many of these compounds
to be simply waste products of metabolism, but they are
now known to possess important functions. One major
category of such compounds is alkaloids. Although
they vary greatly in their chemical structures, alkaloids
have several common characteristics: they possess
nitrogen and alkaline (basic), but none have basic
forms such as quaternary compounds and N-oxides.
Alkaloids are formed as metabolic by-products and
have been reported to be responsible for the
antibacterial activity in most of the medicinal plants
. The alkaloid extracts obtained from medicinal
plant species have multiplicity of host-mediated
antimicrobial, antihyperglycemic, anti-inflammatory
and pharmacological effects -. The quantitative
phytochemical analysis of methanolic extract of
Dendrobium ovatum (L.) for treating liver disorders
could be due to generation of free radicals. The
constituents in selected plant extract alone or in
combinations might be responsible for the observed
hepatoprotective activity. The total phenolic content
was 4.51 mg/g, flavonoid content was 5.34mg/g and
alkaloids content was 48.16 mg/g respectively. The
maximum yield was found to be in alkaloids than in
phenolic and flavonoid contents.
The present study of the phytochemical screening,
determination of alkaloids in different extracts of
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