Preliminary phytochemical screening and Evaluation of Anti ...

Journal of Current Pharmaceutical Research 2010; 3(1): 47-50
JCPR 2010; 2010; 3(1): 47-50
© 2010 Medipoeia
Received: 09/06/2010
Revised: 17/07/2010
Accepted: xx/xx/2010
Preliminary phytochemical screening and Evaluation
of Anti-inflammatory activity of Ethanolic extract of
leaves of Indigofera tinctoria Linn.
Pramod K.Tyagi, Vishnu K. Rai, Ashish K.Pahria, S. Sambath Kumar,
Yogendra Singh, Manoj Sharma and Manoj Goyal
Pramod K.Tyagi , Vishnu K. Rai ,
Ashish K.Pahria, S. Sambath Kumar,
Yogendra Singh and Manoj Sharma
Shri RNS College of Pharmacy, Gormi,
Bhind, M.P
Manoj Goyal
IPS College of Pharmacy Gwalior
(M.P.), India.
ABSTRACT
Objective: The main objective of the present investigation is to preliminary phytochemical
screening and Evaluation of anti-inflammatory activity of Indigofera tinctoria Linn.
Material & methods: in vivo testing of the anti-inflammatory activity of ethanol extract of
leaves of Indigofera tinctoria linn. (EEIT). The ethanol extract of Indigofera tinctoria linn. (250,
500 & 1000 mg/kg bwt, p.o.) Reduced acute paw oedema volume induced by sub-planter
injection of carrageenan (0.1ml of 1% solution) in Wister Albino rats using plethysmometer.
Ibuprofen (100 mg/kg bwt, p.o.) were used as a standard drug (positive controls).
Results: Ethanolic extracts of leaves of Indigofera tinctoria Linn. (500 & 1000 mg/kg bwt)
showed potent anti-inflammatory activity when compared to control as well as positive control
Ibuprofen (standard drug) group. Values are expressed as mean ± S.D. statistical significance was
determined using the student’s t-test. Values with p

Journal of Current Pharmaceutical Research 2010; 3(1): 47-50
been reported in this regard. In the present study, sincere effort has
been attempted to establish the scientific validity of the antiinflammatory
activity of the aerial part of plant.
2. MATERIALS AND METHODS
Plant material
The fresh leaves of the plant Indigofera tinctoria Linn.
were collected from Budhara, Porsa and campus of Shri R.N.S
College of Pharmacy Gormi, Bhind (M.P.). It was further
authenticated by taxonomist Dr. A. N. Rai, department of Botany,
Dr. H.S. Gour University, Sagar,M.P. The leaves were cut into
small pieces and shade dried at room temperature, dried leaves was
subject to size reduction to a coarse powder by using a dry grinder.
Chemicals
Carrageenan was purchased from Himedia Laboratories
Pvt. Ltd., Mumbai and Ibuprofen was obtained from Glenmark
Pharmaceutical Ltd., Mumbai as a gift sample.
Animal experiments
Wister albino rats weighing 200±20 g were taken from
Shri R.N.S College of Pharmacy, Gormi, Bhind (M.P.) and
maintained under constant condition (temperature: 20±2 o C,
humidity: 40-60% , 12-h light/ 12-h dark cycle) for two weeks and
housed in stainless steel cages in a laboratory room. The animals
were access to water ad libitum and standard rodent diet. All
animal experiment conducted in accordance with Institutional
Animal Ethical Committee (IAEC licence
no1030/A/07/CPCSEA.).
Table 1. Phytochemical screening of Petroleum ether & ethanolic
extracts of Indigofera Tinctoria linn.
Preparation of plant Extract
The course powder plant material was extracted with
petroleum ether (60-80 o C) for 72 hours and then using ethanol in
soxhlet extractor for 48 hours (Harbosne et.al., 1984). Alcoholic
extract was concentrated under the vacuum in rotary flash
evaporator and successively in hot air oven till solid to semisolid
mass. Extracts were stored in an airtight container in refrigerator
below 10 o C.
Preliminary phytochemical investigations:-
The petroleum ether extract and ethanolic extract
subjected to preliminary qualitative chemical analysis. Standard
method were used for preliminary phytochemical screening of the
extract was performed to know the phytoconstituents in the extract
.(Kokate. et.al., 2007, Khandelwal et.al.,2004), and it was found
that extract contains carbohydrate, glycosides, proteins, steroid,
amino acids, tannins and flavoniods.
Table 2. Physical contents of leaves of Indigofera Tinctoria linn.
S. NO. PARAMETERS PERCENTAGE
1. Total ash 12.5%
2. Acid insoluble ash 3.5%
3. Water soluble ash 9%
4 Sulphated ash 20%
5. Extractive value 14.2%
6. Loss on drying 6.9%
S.
NO.
PHOTOCHEMICAL
CONSTITUENTS
EXTRACT
PETROLEUM
ETHER
1. Alkaloids - -
2. Fats and oils + -
3. Carbohydrates - +
4. Glycosides + +
5. Steroids + +
6. proteins - +
7. Amino acid - -
8. Tannins - +
9. Flavonoides - +
ETHANOL
Determination of physical constants:-
Leaves analysis parameters like total ash, acid insoluble
ash, water soluble ash, Sulphated ash, loss on drying were use for
the determination of quality and purity of the crude drug and
extractive value were used for evaluation and study of antiinflammatory
activity (Evans et .al., 2002) .
Ethanolic extract of leaves of Indigofera tinctoria linn
(250, 500 & 1000 mg/kg bwt, p.o.) were prepared in tween 80
(1%w/v solution).
Carrageenan induced paw oedema in rats
Pedal inflammation in hind paw of Wister albino rats
(200±20 g) was produced according the method described (by
Winter,et.al., 1962). Oedema was induced by sub-planter injection
(+) represent presence & (-) absence of respective components
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Journal of Current Pharmaceutical Research 2010; 3(1): 47-50
0.1ml of 1% solution of carrageenan into the right hind paw
volume of the rats after the test samples had been administered
orally. The test sample were first dissolved in 1% w/v tween 80
and diluted with saline. The control group received the vehicle,
positive control group received Ibuprofen as reference drug (100
mg/kg, orally). A test solution (250, 500 & 1000 mg/kg bwt,
orally) was administered 1hr prior to injecting carrageenan. Paw
volume were measured up to 3 hr after the carrageenan injection at
interval 60 min, and the volume of the oedema was measured with
a plethysmometer (Rolex pvt ltd, Punjab,India).
Group V treated with 1000 mg/kg bwt of ethanolic extract of
leaves of Indigofera tinctoria Linn. (1% w/v tween-80, orally) as a
test sample.
Statistical analysis:-
Values are expressed as mean ± S.D. statistical significance was
determined using the student’s t-test. Values with p

Journal of Current Pharmaceutical Research 2010; 3(1): 47-50
Leaves analysis parameters (table 2) like as total Ash
(12.5%), acid insoluble Ash (3.5%), water soluble Ash (9%),
sulphated Ash (20%), extractive value (14.2%) and loss on drying
(6.9%).
Ethanolic extracts of leaves of Indigofera tinctoria Linn.
(500 & 1000 mg/kg bwt) showed potent anti-inflammatory activity
when compared to control as well as positive control Ibuprofen
(standard drug) group. The low doses of ethanolic extract (250
mg/kg bwt) did not show inhibition of inflammation, (table-3).
This anti-inflammation activity was dose dependent and found to
be statistically significant. Ethanolic extracts of the dose of 500
and 1000 mg/kg body weight showed inhibition of paw oedema
after 2 hrs. 48.43%, 61% and 3 hrs. 51%, 62% respectively.
The present study indicates that oral administration of
ethanolic extract of Indigofera tinctoria Linn. dose dependently
improve the potent anti-inflammatory activity. The extract lowers
the carrageenan induced rat paw oedema. Further pharmacological
and biochemical investigations are essential to elucidate the
mechanism of action.
ACKNOWLEDGEMENTS
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We are grateful to Shri S.N.Singh Kushwash Chairman
Shri R.N.S. Collage Of Pharmacy Gormi Bhind (M.P.) and Dr. A.
N. Rai, the taxonomist at department of Botany, Dr. H.S. Gour
University, Sagar (M.P), India for supporting to carried out this
project work.
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