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Arif et al. European Journal of Biomedical and Pharmaceutical Sciences
www.ejbps.com
551
PHYTOCHEMICAL AND PHYSICOCHEMICAL PROPERTIES OF HIBISCUS ROSA
SINENSIS LEAVES EXTRACT: A COMPARISON BETWEEN CONVENTIONAL AND
MICROWAVE ASSISTED EXTRACTION
Samreen Fatema1, Ola Basa’ar1, Mazahar Farooqui2 and Pathan Mohd Arif*1
1Post Graduate and Research Center, Maulana Azad College, Aurangabad (MS), India 431001.
2Dr. Rafiq Zakaria College for Women, Navkhanda, Aurangabad (MS) India 431001.
Article Received on 15/05/2018 Article Revised on 05/06/2018 Article Accepted on 25/06/2018
INTRODUCTION
Beautiful flowers of Hibiscus rosa sinensis is well
known in India. It belongs to the Malvacacae and
commonly known as jasvant in India.[1] The leaves of the
plant is known for the treatment of antidiarrhetic and are
antipertensive and antiphlogestic activities.[2] In
ayurvedic system of medicine Hibiscus rosa sinensis is
widely used in India as a demulcent refrigerant drink in
fever and decoction is given in bronchial catarsh.
Sikawar Mukesh and et al[3] also reported the
antihyperlipidemic activity from ethanolic extract of
Hibiscus rosa sinensis. Anti-implantation and
antispermatogenic activities are possess by flowers of
Hibiscus rosa sinensis. Leaves and flowers are also
possess hypoglycemic activity. The petroleum ether
extracts of the leaves and flowers have been shown to
increase the hair growth. Antifungal, insect-repelling and
toxic activities[4] bleeding, soother irritated tissues and
relaxes spasms. It contains anodyne, operient, emollient
and laxatine leaves. The flowers are hermaphrodite and
are pollinated by insects.[5] Flowers of Hibiscus rosa
sinensis are found to be more effective against arterial
hypertension. India intended for hair growth in the
market by the herbal products included various extract of
Hibiscus rosa sinensis Linn. N Adhirajan[6] found the
leaf extract is more effective to increase the length of
hair significantly higher as compare to flower extract.
MATERIAL AND METHODS
Collection of plant leaves
Plant leaves were collected from the local area near to
Maulana Azad College Aurangabad. The leaves were
washed gently and dried under shade. It was grind and
used for the analysis.
Fluorescent test
0.5 gm of samples were added in different solvents and
fluorescent behavior was observed. Normal light
florescent behavior was different in different solvent.
Ash analysis
Accurately weighted 10 gm of sample was taken in
finely clean silica crucible and ignited for 4 hrs with
gradually increasing in temperature up to 300oC. After
ignition of leaves of plant, the residue was remain is
designated as ash. The residue was again ignited with the
interval of 10 min, till to get the constant weight. This
ash was used to determine the three parameters called as
total ash, acid insoluble ash and water soluble ash.
Acid insoluble ash
Acid insoluble ash, ash which is insoluble in dilute HCl.
1 gm of total ash was dissolved in 2 N hydrochloric acid.
Stirred well for the digestion of ash and filtered through
wattman filter paper no. 41. The residue remain after
SJIF Impact Factor 4.918
Research Article
ejbps, 2018, Volume 5, Issue 7, 551-559.
European Journal of Biomedical
AND Pharmaceutical sciences
http://www.ejbps.com
ISSN 2349-8870
Volume: 5
Issue: 7
551-559
Year: 2018
*Corresponding Author: Pathan Mohd Arif
Post Graduate and Research Center, Maulana Azad College, Aurangabad (MS), India 431001.
,
ABSTRACT
The extraction of the herbal drugs are need to be analysed to give the documentary prove of the herbal drugs
against many diseases. Hibiscus rosa sinensis is one of the famous herbal medicine used for the treatment of
bleeding soother irrited tissue and relxes spams. Various extract of Hibiscus rosa sinensis is also used for the hair
growth. Hibiscus rosa sinensis belongs to the Malvaceae family. In the present research work the comparersion of
conventional extraction (CE) and microwave assisted extraction (MAE) was done. It is observed that both are
hydro extract but gives different results for phytochemicals and physicochemical properties. Carbohydrate,
glycosides proteins, amino acid, phenolic compounds and tannins were present in both the extracts. The CE is
found to be active against E.coli, B.Subtilis, S.Aureus bacteria while MAE is found to be inactive.
KEYWORDS: Phytochemical, physicochemical, microwave extract, aqueous extract, antibacterial, ant-
tuberculosis, anti-malarial.
Arif et al. European Journal of Biomedical and Pharmaceutical Sciences
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552
filtration is ignited in clean silica crucible by gradually
increase in temperature up to 300oC. The residue was
cooled and weighted and again kept for ignition till to get
the constant weight. The residue is remain after ignition
is acid insoluble ash. The percentage was calculated for
the acid insoluble ash.
Water soluble ash
1 gm of total ash was boiled with20ml of double distilled
water. The residue was collected by filtered through
wattman filter paper no 41. Residue was washed with hot
water and kept for ignition not more than 400oC. The
weight of residue was subtracted from total ash. This
difference between residue and total ash represent the
water soluble ash. The percentage was calculated.
Bulk density
Bulk density and tab density were determined by
densitometer (Bio Technics India, serial No. 40195,
Model BTI-09). 50 cm3 of powder was introduced into
the 50 ml graduated cylinder. The dropping interval of
the cylinder was two sec at the height of 2 cm three times
on the hard wooden surface. The bulk density was
calculated by dividing the weight of the sample in grams
by the final volume in cm3 of the sample contained in the
cylinder.
Tapped density
50cm3 of powder was introduced into the 50 ml
graduated cylinder. The dropping interval of the cylinder
was two sec at the height of 2 cm 100 times on the hard
wooden surface. The tab density was calculated by
dividing the weight of the sample in grams by the final
volume in cm3 of the sample contained in the cylinder.
The compressibility of the powder was evaluated using
the HR (Housner Ratio). The Housner ratio may be
defined as ratio of tap density and bulk density.
HR=
Carr’s Index (Compressibility Index- CI): This was
calculated by using the formula:
CI = X 100
Extraction procedure
Conventional Extraction
Accurately weight 30 gms of sample was introduced into
the 500 ml round bottom flask (which was first clean by
very dilute hydrochloric acid and then distilled water)
with 300 ml double distilled water. The pourciline pieces
were add to avoid bumping of the sample. The condenser
was fitted with circulation of water. The sample was
refluxed on flame for six hrs. The sample was cooled and
filtered by the suction pump. The excessive water was
evaporated for the preservation of the sample and it was
kept at 4oC for 12 hrs. The percentage of the extract was
calculated.
Microwave Assisted Extraction
30 gms of the sample was kept in the clean round bottom
flask. 300ml of double distilled water was used as the
solvent. The porciline pieces were added to avoid
bumping of the sample. The condenser was fitted with
circulation of water. The sample was refluxed by
microwave radiations using microwave oven (Catalyst
microwave synthesizer Sr. No. 130602954) for 30 min at
60% power, 420 watt and 120oC. The sample was cooled
and filtered by the suction pump. The excessive water
was evaporated for the preservation of the sample and it
was kept at 4oC for 12 hrs. The percentage of the extract
was calculated.
Physicochemical test
Physicochemical parameters like relative density,
viscosity, surface tension and refractive index were
measured of the solutions of different ppm.
Relative density
Clean and dry empty density bottle with stopper
weighted accurately. The density bottle was filled with
double distilled water up to it fall from the bottle and
stopper was fitted and the bottle was cleaned from
outside. The bottle was weighted.
The procedure was repeated for the samples. The density
was measured by taking difference between bottle with
sample and empty bottle. Relative density was calculated
by the formula,
Relative density =
Where ρ1 is density of the water and ρ2 is the density of
the sample.
Viscosity
The different concentration in ppm of the samples were
prepared by using double distilled water. The Ostwald’s
viscometer was cleaned by NaOH to remove greasy
impurities than with chromic acid and finally with the
distilled water. The 10 ml of double distilled water was
inserted in viscometer from large diameter tube. And the
sample was sucked through second tube of the same
viscometer till it rises with 2-3 cm above the mark. By
keeping stop watch ready the liquid was allowed to
decent down the time required to flow of the liquid
between two points was noted. The same procedure was
repeated for the samples which have to study.
Viscosity was measured by sing formula,
η2=
Surface tension
The different concentration in ppm of the samples were
prepared by using double distilled water. The
stalagmometer was cleaned by NaOH to remove greasy
impurities than with chromic acid and finally with the
distilled water. The rubber tubing with the with a screw
Arif et al. European Journal of Biomedical and Pharmaceutical Sciences
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clip was attached to the top of the stalgmometer. The
flat end of the stalagmometer was dipped into the
standard solution (double distilled water) suck through
the water tubing until the water level rises above the
mark. The screw was adjust the pressure until the rate of
the drop was 10 to 15 per minute. The number of drops
were counted for double distilled water when passes
from upper mark to the lower mark.
The stalagmometer was removed and rinse with alcohol
and dried. Stalagmometer was filled with the test sample
and number of drops were determine. Same procedure
was repeated for every concentration three times and
mean was taken.
The surface tension was than calculated by the formula,
Where γ1 and γ2 are the surface tension of the double
distilled water and the sample respectively.
And n1, ρ1 and n2, ρ2 are the number of drops and relative
densities of the double distilled water and samples under
study respectively.
Refractive index
Refractive index depends upon temperature and
concentration. However the specific refraction is
independent of temperature and it is characteristic
property of the liquid. The refractive index was measured
by Abbe’s refractometer. The refractometer was placed
on the table near to the window so that sufficient light
should reached to the prism. The prism box was opened
b turning the lock nut and both the phases of the prism
was cleaned with the help of cotton wool by the acetone
and prism box was closed after drying. Few drops of the
liquid were pumped through the small hole on the prism
box with the help of the dropper. The crosswire of the
telescope was focus by rotating the eye piece and the
mirror was adjust for the reflection of maximum light
towards the prism box. The prism box was moved
forward and backward until the clear boundary between
the light and dark region was appear. The scale reading
was noted down. And refractive index were calculated by
using the formula,
Specific refraction (r) =
Where ρ is the density of the samples.
Qualitative Phytochemical Screening
The different qualitative chemical tests were performed
for establishing profile of given extract for its chemical
composition. Qualitative Phytochemical analysis was
done using the following procedures.
Detection of alkaloids
Solvent free extract, 50 mg was stirred with 5 mL of
dilute hydrochloric acid and filtered. The filtrate was
tested carefully with various alkaloidal reagents as
follows.
i) Mayer’s test: To a few mL of filtrate, two drop of
Mayer’s reagent added by the side of test tube. A white
or creamy precipitate indicated the test as positive.
Mayer’s reagent: Mercuric chloride (1.36 g) was
dissolved in 60 mL of water and potassium iodide (5.0 g)
was dissolved in 10 mL of water. The two solutions were
mixed and made up to 100 mL with water.
ii) Wagner’s test: To a few ml of filtrate, few drops of
Wager’s reagents were added by the side of the test tube.
A reddish-brown precipitate confirmed the test as
positive.
Wagner’s reagent
Iodine (1.27 g) and potassium iodide (2 g) was dissolved
in 5 ml of water and made up to the 100 ml with distilled
water.
iii) Hager’s test: To a few ml of filtrate 1 or 2 of
Hager’s reagent (saturated aqueous solution of picric
acid) were added. A prominent yellow precipitate
indicated the test as positive.
Detection of Carbohydrate
The extract 100 mg was dissolved in 5 ml of water and
filtered. The filtrate was subjected to the following tests.
i) Molisch’s tests: To 2 ml of filtrate, two drops of
alcoholic solution of α-naphthol were added, the
mixture was shaken well and 1 ml of concentrated
sulphuric acid was added slowly along the sides of
test tube and allwed to stand. A violet ring indicates
the presence of carbohydrates.
ii) Fehling’s test: One ml of filtrate was boiled on
water bath with 1 ml each Fehling’s solutions A and
B. Red precipitate indicates the presence of suger.
Fehling’s solution
Solution A; copper sulphate (34.66 g) was dissolved in
distilled water made up to 500 ml with distilled water.
Solution B: potassium sodium tartarate (173 g) and
sodium hydroxide (50 g) was dissolved in water and
made up to 500 ml.
iii) Benedict’s test: To 0.5 ml of filtrate, 0.5 ml of
Benedict’s reagent was added. The mixture was
heated on a boiling water bath for 2 min. A
characteristics colored precipitate indicates the
presence of sugar.
Benedict’s reagent
Sodium citrate (173 g) and sodium carbonate (100 g)
were dissolved in 800 ml distilled water and boiled to
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make it clear solution. Copper sulphate (17.3 g)
dissolved in 100 ml distilled water.
iv) Barfoed’s test: To 1 ml of filtrate, 1 ml of
Barfoed’s reagent was added and heated on a boiling
water bath for 2 min. red precipitate indicates the
presence of suger.
Barfoed’s reagents: copper acetate 30.5 g was dissolved
in 1.8 ml of glacial acetic acid.
Detection of glycosides
For the detection of glycosides, 50 mg of extract was
hydrolysed with concentrated hydrochloric acid for 2 hrs
on water bath, filtered and the hydrolysate was subjected
to the following test.
i) Borntrager’s test: To 2 ml of the filtrate
hydrolysate, 3 ml of chloroform was added and
shaken, chloroform layer was seprated and 10%
ammonia solution was added to it. Pink color
indicates the presence of glycosides.
ii) Legal’s test: Fifty mg of extract was dissolved in
pyridine, sodium nitroprusside solution was added
and made alkaline using 10% sodium hydroxide.
Presence of glycoside was indicated the pink color.
Detection of Saponins by Foam test
The 50 mg was diluted with distilled water and made up
to 20 mL. The suspension was shaken in a graduated
cylinder for 15 min. A two cm layer of foam indicated
the presence of saponins.
Detection of proteins and Amino acids
The extract (100 mg) was dissolved in 10 ml of distilled
water and filtered through Whatman filter paper no. 41
and filtrate was subjected to test of proteins and amino
acids.
i) Millon’s test: To 2 mL of filtrate, few drops of
Millon’s reagent were added. A white precipitate
indicated the presence of proteins.
Millon’s reagent
Mercury (1 g) was dissolved in 9 mL of fuming nitric
acid. When the reaction was completed, equal volume of
distilled water was added.
ii) Biuret test: An aliqot of 2 ml of filtrate was treated
withone drop of 2% copper sulphate solution. To
this 1 ml of ethanol (95%) was added, followed by
excess of potassium hydroxide pellets. Pink color in
the ethanolic layer indicates the presence of proteins.
iii) Ninhydrine test: two drops of ninhydrine solution
(10 mg of ninhydrine in 200 ml of acetone) were
added to two ml of aqueous filtrate. A characteristic
purple color indicates the presence of amino acids.
Detection of phenolic compounds and tannins
i) Ferric chloride test: The extract (50 mg) was
dissolved in 5 ml distilled water. To this few drops
of neutral 5% ferric chloride solutions were added.
A dark green color indicates the presence of
phenolic compounds.
ii) Gelatin test: The extract (50 mg) was dissolved in 5
ml of distilled water and 2 ml of 1% solutions of
gelatin containing 10% sodium chloride was added
to it. White precipitate indicates the presence of
tannins.
iii) Lead acetate tests: the extract (50 mg) was
dissolved in distilled water and 3 ml of 10% lead
acetate solution was added. A bulky white
precipitate indicates the presence of flavonoids
compounds.
iv) Alkaline reagent test: an aqueous solution of the
extract was treated with the 10% ammonium
hydroxide solution. Yellow fluorescence indicates
the presence of flavonoids.
Spectral analysis: UV-Visible
The powder of extract was dried and dissolved in
distilled water to prepared solution of approximately 50
ppm. The spectra was recorded in the range from 190 to
800 nm by using double beam spectrophotometer of
Model Elico-159 and λmax is determine from the
software Spectra treat.
IR Spectra
The FTIR instrument IRT3000, JASCO, having serial
no. B051061016, and the spectra were recorded using
spectra manager. IR instrument is calibrated by using
polystyrene. Spectra were recorded by using potassium
bromide (KBr) of IR grade manufactured by Marck life
sciences. KBr was kept in hot oven at 50oC for half hour
to free it from moisture. Spectra of that dry KBr is
measured within IR range. The samples of leave extract
were crushed to make it fine powder and mixed with dry
KBr, and spectra of the samples were measured.
Anti-microbial study
Antibacterial activity was investigated using disc
method. With the help of sterile wire loop, the test was
inoculated into a test tube containing Mueller Hinton
broth. The O.D of the inoculums was adjusted in
between 0.08-0.1.
As per the composition, Mueller Hinton Agar was
prepared by using sterile distilled water and was
sterilized at 121oC at 15lb pressure for 15 min in an
autoclave. The medium was cooled at room temperature
and poured in sterile petri plates and were allowed to
solidify. Bacterial inoculums was swabbed over the
medium using sterile cotton swab. Sterile disc was
placed on medium, on which 20 µl of complex
suspension was added. Zone of inhibition were observed
and measured after incubation at 30oC for 18-20 hrs.
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RESULT AND DISCUSSION
Fluorescent test
The powdered samples were treated with different
chemicals and observed with naked eyes. The results
were summarized in table 1. It is observed that the dark
green color of powder is retaining its color in benzene,
faint in presence of butanol, ethanol, chloroform, acetic
acid etc. It becomes blackish brown in strong inorganic
acids or base. This can attributed to oxidation or
reduction or incomplete decomposition of compounds
and formation of carbon particles.
Table 1: Florescent test for leave powder.
Sr. No.
Solutions
Observation
1
Powder as such (P)
Dark green
2
P + n-butanol
Whitish green
3
P + Conc. HCl
Brown
4
P + Conc. HNO3
Reddish orange
5
P + Conc. H2SO4
Blackish brown
6
P + Chloroform
Light whitish green
7
P + Ethanol
Whitish green
8
P + Glacial acetic acid
Light whitish green
9
P + 1N HCl
Almond caream
10
P +1N NaOH
Yellowish green
11
P + 5% HCl
Cream
12
P + 5% NaOH
Chocaloty
13
P + benzene
Dark green
Ash value
The total ash content of the powder is formed to be 6.3%.
The acid insoluble ash was found to be 39%. Whereas
water soluble as 18%. The ash is mostly consist of
transition elements i.e. cobalt or magnesium. The tapped
density and bulk density of the powdered material is
found to be 0.5535 and 0.4169 respectively. Carr’s index
and housner’s ratio is found to be 24.68% and 1.3277
respectively. B.V Basavaraj et al[7] also report the bulk
density, tapped density, Carr’s index and housner’s ratio
of Hibiscus rosasinensis Linn, from that they get the
result as 0.42, 0.44, 4.54% and 1.047 respectively.
The results of present study are given in table 2.
Table 2: Ash analysis and densities of leave powder.
Sr. No.
Ash
Results
1.
Total Ash
6.3 %
2.
Acid Insoluble
39 %
3.
Water Soluble
18 %
4.
Bulk density
0.4169
5.
Tapped density
0.5535
6.
Housner Ratio
1.3277
7.
Carr’s index
24.68%
Percent Extraction
The extractive values are given in table 3. It is less when
conventional method is used. When the extraction is
carried out by microwaves, the extractive value is more
as well as the time of extraction required is also less.
Mehran Moradalizadeh et al[8] report that percent
extraction of microwave assisted hydro extraction
(MAHE) and conventional hydro extraction (CHE) was
compared and it was found that MAHE gives more
percentage as compared to CHE with significantly
shorter time of extraction (15 min against 3 hrs for hydro
Extraction). Microwave assisted extraction has many
advantages such as shorter extraction time, lesser solvent
consumption, higher extraction rate and better products
with lower cost.[9,10]
Table 3: Extractive value of Hibiscus rosa sinensis plant leaves.
Sr. No.
Solvent
Percentage
1.
Conventional Hydro Extraction
12.26
2.
Microwave Assisted Hydro Extraction
15.16
Physico-chemical properties
Physicochemical parameters determine were Relative
density, viscosity, surface tension, Refractive index in
table 4 and 5. The relative density and surface tension of
CE was found to be higher. The refractive index was
found to be less in MAE. The surface tension for the
MAE found to be approximately constant over the range
of concentration (fig1 to 4). The extract contains
saponins which are surface active agents. They may get
disperse into water molecules, reducing the surface
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tension of the solutions. Sometimes these molecules may
get aggragate to form micelle. The polar group present in
the molecules may get oriented at particular
concentration, known as critical micelle concentration
(CMC). Such decrease in surface tension is reported in
the form 72 mNm-1 (pure water) to 25mNm-1 for extract
of Pfaffia glomcrata and Hebanthe erintha plant
extract.[11] There are different complex macromolecules
present in the plant extract. Some of them like
polysaccharides are rich in hydroxyl groups. Hence they
bind with each other and take up water and become rich
in cis-OH group. They allow aggregation from chain to
chain via hydrogen bonding. The hydration becomes
more complicated due to the interchange cross linking.[12]
Hence hydration affects the physic-chemical properties
of extract.
Table 4: Physicochemical properties of CE of Hibiscus rosa sinensis Leaves.
Sr.
No.
Solution in ppm
Relative
density
Viscosity
(Pascal sec)
Surface tension
(Newton/meter)
Refractive
index
1.
5
1.00192
0.83739
69.8346
0.99556
2.
10
0.99697
0.83326
67.0941
0.99898
3.
20
0.99849
0.83447
60.8967
0.99897
4.
40
0.99828
0.83436
62.8473
0.99919
5.
60
0.99707
0.83334
60.8101
1.00040
6.
80
0.99919
0.83512
60.9393
1.00056
7.
100
1.00282
0.83814
61.1610
0.99466
Table 5: Physicochemical properties of MAE of Hibiscus rosa sinensis Leaves.
Sr.
No.
Solution in ppm
Relative
density
Viscosity
(Pascal sec)
Surface tension
(Newton/meter)
Refractive
index
1.
5
0.9988
0.9029
57.2031
0.9997
2.
10
1.0019
0.9057
60.9731
0.9947
3.
20
1.0032
0.8720
63.0109
0.9953
4.
40
0.9990
0.8684
57.2314
0.9995
5.
60
1.0002
0.8694
59.0553
0.9983
6.
80
1.0007
0.8698
57.3514
0.9978
7.
100
1.0009
0.8739
57.6200
0.9976
020 40 60 80 100
0.997
0.998
0.999
1.000
1.001
1.002
1.003
1.004
Relative Density (Dynes/cm3)
concentration (ppm)
CHE
MAHE
020 40 60 80 100
0.83
0.84
0.85
0.86
0.87
0.88
0.89
0.90
0.91
Viscosity (Pascal sec)
concentration (ppm)
CHE
MAHE
Fig. 2: Relative density of CE and MAE. Fig. 1: Viscosity of CE and MAE.
020 40 60 80 100
56
58
60
62
64
66
68
70
Surface tension (Newtons/ meter)
concentration (ppm)
CHE
MAHE
020 40 60 80 100
0.994
0.995
0.996
0.997
0.998
0.999
1.000
1.001
Refractive ondex
concentration (ppm)
CHE
MAHE
Fig. 3: Surface tension of CE and MA. Fig. 4: Refractive index of CE and MAE.
Arif et al. European Journal of Biomedical and Pharmaceutical Sciences
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557
Phytochemical test
Both extract shows almost identical Phytochemicals
(table 6). The alkaloid gives negative test, except
wagner’s test in MAHE. The carbohydrate test positive,
foam test also positive indicating presence of saponins.
The proteins, amino acids, phenolic compounds and
tannins are present in both the extracts. Faten R and et al
found that 70% ethanol extract give positive test for
carbohydrate glycosides, steroids/ terpenoids, flavonoids
and tannins.[13] Uzama Danlami et al[14] shows that
hexane wxtract of securinega virosa leaf shows
maximum positive results for phytochemcal like
saponins, alkaloids, flavonoids etc.
Table 6: Qualitative test for Hibiscus rosa sinensis leaves extract.
Sr. No.
Reagent
CE
MAE
1.
Detection of Alkaloids
A.
Mayer’s test
-ve
-ve
B.
Wagner’s test
-ve
+ve
C.
Hager’s test
-ve
-ve
2.
Detection of carbohydrate
A.
Molish test
+ve
+ve
B.
Fehling’s test
+ve
+ve
C.
Benedic test
-ve
-ve
D.
Barfoad’s test
+ve
+ve
3.
Detection of Glycosides
A.
Borntrager’s test
-ve
-ve
B.
Legal’s test
-ve
-ve
4.
Test for saponins
+ve
+ve
5.
Detection of proteins and amino acid
A.
Millon’s test
-ve
-ve
B.
Nitric acid test
+ve
+ve
C.
Biuret test
+ve
+ve
D.
Ninhydrine test
+ve
+ve
6.
Detection of phenolic compound and tannins
A.
Ferricchlorid test
+ve
+ve
B.
Gelatin test
-ve
-ve
C.
Lead acetate test
+ve
-ve
D.
Alkaline reagent test
+ve
+ve
FTIR spectrum
The IR spectrum of extract of Hibiscus rosa sinensis was
recorded from FTIR instrument IRT3000, JASCO,
having serial no. B051061016. (fig5) Though it contains
a mixture of compounds but still in order to find out
various functional groups and a general finger print of
samples, it will help. There are various IR bands
observed. Which are represented in table 7.
Table 7: IR bands of Hibiscus rosa sinensis plant leaves extract.
Band (cm-1)
Intensity
Functional group
3382
Very broad
Inter molecular hydrogen bonding
1583
Sharp
Aldehydic ketone
1417
Sharp
C-H bending vibrations
1121
Broad
C-O-C asymmetrical stretching
770
Broad
Four adjacent hydrogen
657
Broad
Aromatic hydrocarbon
604
Broad
C=C bend
Arif et al. European Journal of Biomedical and Pharmaceutical Sciences
www.ejbps.com
558
Fig. 5: FTIR spectrum of Hibiscus rosa sinensis leaves extract.
UV spectrum
The UV spectra of both the extract shows λmax at 196
nm and 206nm for conventional and microwave assisted
extract respectively. The spectral trend in conventional
and microwave assisted extract is somewhat similar (Fig
6 & 7).
Fig. 6: UV-visible spectra of CE. Fig. 7: UV-visible spectra of MAE.
Antibacterial study
The extracts are screened for the biological activity
particularly for antibacterial activity. We observed that
MAE is totally inactive against all the tested bacteria
table 8. In MAE, sudden energy is supplied to the
molecule, due to that some molecule may get
decomposed by the heat generated inside the vessel.
Table 8: Antibacterial, anti tuberculosis and anti malarial properties of Leasves extract.
Sr. No.
Bacteria
MAE
CE
1.
E. coli
- -
Active
2.
B. Subtillis
- -
Active
3.
S. Typhi
- -
Inactive
4.
S. Aureus
- -
Active
5.
T.B
- -
-----
6.
Maleria
--
-----
CONCLUSION
Although the extractive value of microwave assisted
hydro extraction is higher but the Phytochemical
ingrideint and physicochemical properties differs
compared to conventional hydro extraction. Even the
biological activity is also changes with the type of
extraction methods.
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