Pharmacognostical & Phyto-Chemical Investigation of Cissus Quadrangularis Linn. Stem PDF

Publication Ref No.
: IJPRD/2011/PUB/ARTI/VOV-3/ISSUE-1/MARCH/020
ISSN 0974 9446
PHARMACOGNOSTICAL & PHYTO-CHEMICAL INVESTIGATION OF CISSUS QUADRANGULARIS

LINN. STEM
Anuj K. Srivastava1*, Priyanka Srivastava1,
B. R. Behera2, A. K. Shrivastava3
1
Anuj Srivastava
School of Pharmacy, Suresh Gyan Vihar University,

Jaipur (Raj.), India
2
The Pharmaceutical College Ingipali,
Bargarh (Orissa), India
3
Anand Colleges of Pharmacy, Agra, (U.P.), India
Email:anuj_pharmacognosy@rediffmail.com
ABSTRACT
Cissus quadrangularis (Family-Vitaceae) is found in hotter parts of India and neibouring countries
Pakistan, Sri Lanka, and Malaysia .Stem of the plant are used as anthelmintic, laxative, chronic ulcer,
,analgesics and in Bone healing activity. The Stem of Cissus quadrangularis was collected and
powdered. Ethyl acetate extract was prepared by using Cold Maceration and subsequently fractioned
with hydro alcoholic (50:50) for detailed chemical analysis. Fluorescence characters of different
Macerated extract and powder were noted under ultra violet light and under normal ordinary light, which
signifies their characteristics. The total ash value, acid insoluble ash value and water soluble ash value
were 12.5%, 2.5% & 10.0%, respectively. The percentage yields for ethyl acetate, Hydro alcoholic were
2% and 1.8%. respectively. Reaction behaviors of powder stem of Cissus quadrangularis with 10
different reagents were also noted, which revealed characteristic development of particular color.
Preliminary qualitative chemical tests of extracts were found positive results for Phytosterol, flavonoids
and triterpenoids in Ethyl acetate fraction, Hydro alcoholic fraction was positive to carbohydrate, tannin
and amino acid and vitamin C. Flavonoids and triterpenoids are the active constituents in stem of
Cissus quadrangularis and may be responsible for its pharmacological activities. The results of the
study could be useful in setting some diagnostic indices for the identification and preparation of a
monograph of the plant.
Key Words: Cissus quadrangularis, Maceration, Physicochemical analysis
International Journal of Pharma Research and Development Online
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Publication Ref No.: IJPRD/2011/PUB/ARTI/VOV-3/ISSUE-1/MARCH/020
INTRODUCTION - MATERIALS & METHODS

Plant material collection and authentication:
The mature green stem of Cissus
quadrangularis were collected locally from
Fatehpur District, Uttar Pradesh, India. In the
month of June 2009.The plant herbarium was
prepared, authentified by deposited in the
department of Botany, University of Rajasthan,
Jaipur.
(A voucher specimen No. RUBL-20615).
Drying and size reduction of Stem
The stems of Cissus quadrangularis were
subjected to shade drying and further crushed
to powder, and then the powder is passed
through the mesh 22 and stored in air tight
container for further use.
Pharmacognostical studies
Macroscopic
Morphological studies of Cissus quadrangularis
were done. The Size, shape, taste, colour and
odour were determined. (Table 1&2) (Figure 1)
Microscopic
Microscopic studies were done by preparing a
thin hand section of Cissus quadrangularis
stem. The section was cleared with chloral
hydrate solution, stained with phloroglucinol and
hydrochloric acid, and mounted with glycerin. A
separate section was prepared and stained with
iodine solution for the identification of starch
grains. Microscopic studies of stem showed
epidermis single layered, covered externally
with thick cuticle; epidermal cells thin-walled,
cork and single layered cork cambium; four
patches of Collenchymatous cells present in
cortical region, Collenchymatous vascular
bundles capped by sc1erenchymatous sheath
cambium, idioblasts containing raphides and
isolated acicular crystals of calcium oxalate
present in the outer region of cortex.[5] Powder
of the dried leaves was used for the observation
of powder microscopical characters. The
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powered drug was separately treated with

phloroglucinol and HCL solution, glycerin and
iodine to determine the presence of Beaded
epidermal
cell,
Sclariform
vessels,
Collenchymas Phloem fiber, Raphides and
Acicular crystals of calcium oxalate, Fibres ,
trichomes and starch grains [6].(Table 2) (Figure
2)
Determination
of
Physicochemical
Parameters
Powder of stem of Cissus quadrangularis were
subjected
to
following
physicochemical
parameters. Loss on drying, Total Ash value,
acid insoluble, water soluble, sulphated ash
value, Ethanol soluble, water soluble and
Petroleum ether soluble extractives and PH
determination. The results of physicochemical
parameters are reported [7]. (Table 3)
Treatment of the powder drug with different
chemical reagents
The powdered drug was subjected to various
chemical reagents and noted under ultraviolet
and under normal ordinary light, which signifies
their characteristics. Behavior of Cissus
quadrangularis stem with different chemical
reagent was tabulated in Table 4 (i). [8]
Fluorescence analysis of powder drugs
Fluorescence characters of powdered drug
were noted under ultraviolet and under normal
ordinary light, which signifies their
characteristics[11], Results are displaced inTable
4 (ii).[9]
Extraction of Cissus quadrangularis stem
The shade dried and 22 mesh powdered stem
of Cissus quadrangularis were subjected to
successive cold extraction with Ethyl acetate
and hydro alcoholic (50:50) solvents so as to
get respective 2.0 % w/w and 2.2 % w/w
extracts after due filtration and evaporation to
dryness and stored in a desiccators over
anhydrous Calcium hydroxide for further use.
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Preliminary phytochemical screening of

powdered stem
Test for carbohydrates, gums and mucilage,
proteins and amino acids, alkaloids, glycosides,
phytosterol, Fixed oil & Fats[10], flavonoids,
tannin and phenolic compounds, saponins and
volatile oil were carried out, Results are
displaced in table 5.[11].
RESULTS AND DISCUSSION
The Morphological studies revealed that the
stem is in Green color with Characteristic odour
and slightly characteristic taste. The Cissus
quadrangularis stems are a varying lengths;
stern quadrangular, 4-winged, internodes
constricted at nodes; internodes 4-15 cm long
and 1-2 cm thick. Microscopic studies of stem
showed epidermis single layered, covered
externally with thick cuticle; epidermal cells thinwalled, cork and single layered cork cambium;
four patches of Collenchymatous cells present
in cortical region, Collenchymatous vascular
bundles capped by sc1erenchymatous sheath
cambium, idioblasts containing raphides and
isolated acicular crystals of calcium oxalate
present in the outer region of cortex and also in
a number of cells throughout the region; starch
grains present. Powder study reveals the
presence of covering trichome, Beaded
epidermal
cell,
Sclariform
vessels,
Collenchymas Phloem fiber, Raphides and
Acicular Crystals of calcium oxalate, Fibres,
Pitted Parenchymatous cells. The total ash
value, acid insoluble ash value, water soluble
ash and sulphated ash values were observed to
be 12.5%, 2.5%, 10.0% and 5.0% respectively.
The Ethanol soluble, water soluble and
Petroleum ether soluble extractives were
observed to be 3.0%, 18.0%, & 2.0%.
respectively. Behaviour of powder of stem of
Cissus quadrangularis with different chemical
reagents was also observed. It exhibited Brown,
yellow, yellowish brown, yellow, light brown
colours with Con. HCL, Con. HNO3, 5% KOH,
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Saturated picric acid solution and ammonia

solution respectively. The quantitative chemical
test reveals the presence of carbohydrate;
protein
and
amino
acids,
phytosterol,
flavonoids, tannin & phenolic compounds, gum
mucilage and saponins. The active constituents
like phytosterol, flavonoids and triterpenoids of
stem of Cissus quadrangularis are responsible
for pharmacological activities.
The aim of the specific targeting is to enhance
the efficiency of drug delivery and at the same
time to reduce the toxicity and side effects. A
number of novel drug delivery systems have
emerged encompassing various routes of
administration, to achieve controlled and
targeted drug delivery, magnetic micro carriers
being one of them. These micro carriers
include.
ACKNOWLEDGEMENT
The author is thankful to the Chanceller, Suresh
Gyan Vihar University, Jaipur, And Principal of
Gyan Vihar School of pharmacy, Jagatpura
Jaipur-302025
Rajasthan
for
providing
laboratory and other facilities.
REFERENCES
1. Herb collectors and growers of Chhattisgarh,
India associated with medicinal herb Hadjod
(Cissus
quadrangularis),
http://botanical.com/site/column_poudhia/43_ha
djod.html.
2. Rajpal V. Standardization of Botanical
Testing and Extraction Methods of Medicinal
herbs. Vol.1, Eastern Publishers, New Delhi
2008, p.77.
3. Pankaj O., the traditional healers and natives
of Bhopalpatnam region, Chhattisgarh, India
having rich traditional medicinal knowledge
about common herbs insects and other animals
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209
http://botanical.com/site/column_poudhia/121_b
hopalpatnam.html.
4. Kokate, C.K., Practical Pharmacognosy, 4th
edn, Vallabh Prakashan, Delhi, 1994, pp 15-29
5. Ayurvedic Pharmacopoeia of India, 1986.
The Ministry of Health and Family Welfare.
Govt. of India, New Delhi, Vol III, Third edition,
pp. 21-22.
6.
Khandelwal, K.R., 2005. Practical
Pharmacognosy,
fourth
edition,
Vallabh
Prakashan, New Delhi; p.p. 145.
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7. Indian Pharmacopoiea, Vol. II, 3rd edn.

Ministry of health and family welfare, Govt. of
India,
Controller of publications, New Delhi
1985, p. A-53.
8. Vidhu R., et al. 2007. Hamdard Medicus 50,
109-114.
10 Mahajan R., Practical Biochemistry (Lab.
Manual) For Student, 1st edn, Vayu education
of India 2009.
11. Rao D.G., A Manual of Practical
Biochemistry, 1st edn, Birla Prakashan, Delhi
2006,p.16.
TABLES & FIGURES

Table 1: Identification of Morphological features of Cissus quadrangularis:
Leaf
:
leaves simple or lobbed, sometimes 3-folialate, dentate
Stem
Flower
Fruit
Table 2:
stem of varying lengths; stern quadrangular, 4-winged, internodes

constricted at nodes; internodes 4-15 cm long and 1-2 cm thick; buff
colored with greenish tinge, angular portion reddish-brown.
:
Flowers bisexual, tetramerous, in umbellate cymes, opposite to the leaves,
Calyx cup-shaped, obscurely 4-lobed.
:
Fruit globose or obovoid fleshy berries, one seeded, dark purple to black;
seeds ellipsoid or pyriform. Flowering and fruiting time May-June.
Powder Microscopy
S. No.
1
2
3
4
Microscopic
5
6
7
8
9
10
11
Features
Nature
Colour
Odour
Taste
Observation
Coarse powder
Cream colour light yellow
Slightly characteristic
Slightly characteristic
Beaded epidermal cell

Sclariform vessels
Present
Present
Collenchymas cell
Phloem fiber
Raphides and Acicular
Crystals of calcium oxalate
Fibres
Pitted Parenchymatous cells
Present
Present
Present
Present
Present
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Table 3:
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Results of Physico-chemical parameters

S .No.
Parameter
1
Loss on drying
2
Values (%) w/w

2.5
Total ash
12.5
Acid insoluble ash
2.5
Water soluble ash
10.0
Sulphated ash
5.0
Extractive values
Water soluble extractive
18.0
Alcohol soluble extractive
3.0
Petroleum ether extractive
2.0
Table 4: Quantitative Chemical evaluation

4 (i) Powder analysis with Chemical agents
Reagent
Colour observed
Powder as such
Whitish yellow
Powder + conc. HCl
Brown
Powder + conc. HNO3
Yellow
Powder + conc. H2SO4
Brown
Powder + Glacial acetic acid
Light yellowish white
Powder + 5% NaOH
Brown
Powder + 5% KOH
Yellowish brown
Powder + 5% ferric chloride
Light yellow
Powder + picric acid(Saturated solution)
Yellow
Powder + ammonia
Light brown
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4 (ii) Fluorescence analysis of powder drugs

Fluorescence Observed
Chemicals/Reagents
Short Wavelength
Powder as such
Powder + 1N NaOH IN
Methanol
Powder + 1N NaOH in
water
Powder + 50% HCl
Powder + 50% HNO3
Powder + 50% H2SO4
Powder + Petroleum
ether
Powder + Chloroform
Powder + picric acid
Powder + 5% ferric
chloride solution
Powder + 5% iodine
Powder + methanol
Powder+ HNO3+NH3
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Long Wavelength
Cream
Brown
Brown
Reddish brown
Green
Green
Dark green
Light brown
Reddish brown
Brown
Brown
Dark brown
colorless
Yellow
Green
Brown
Yellowish brown
Reddish brown
Greenish brown
Colorless
Yellowish green
Black
Light brown
Reddish brown
Table 5: Phytochemical analysis of powder and Stem Extracts of Cissus quadrangularis

Chemical Test/Reagent
Powder
Drug
Ethyl
acetate
Extract
Hydro
alcoholic
Extract
Test for Carbohydrate

Molischs Test
Fehlings Test
Benedicts Test
Barfoeds Test
Test for starch
Test for Gums and Mucilages
+
+
+
+
+
+
+
+
+
+
+
Test for Proteins and Amino acids

Ninhydrin test
Biuret Test
Millans Test
+
+
+
+
+
Xanthoprotic Test
Test with tannic acid
Test with heavy metals
Test for Fixed oils and Fats

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Spot Test
Saponofication Test
Mayers reagent test
Dragondraffs reagent Test
Wagners reagent Test
Hagers reagent test
Tannic acid test
Legals test
Balgets test
Borntragers test
Test for cardiac glycosides

(Keller-killiani test )
Test for Cyanogenetic glycosides
Liebermanns Test
+++
Liebermanns burchards Test
++
++
Salkowski Test
+++
Ferric chloride test
++
+++
Fluorescence Test
++
+++
Reaction with alkali and acid
Lead acetate test
+++
Shinoda Test
++
+++
++
++
Test For Alkaloids
Test for Glycosides
Test for Phytosterol
Test for Flavonoids
Test for Tannins and phenolic compounds

Test with Salt of Copper and Lead
+
acetate
Test with Ferric chloride Solution
+
Test for Saponins
Foam test
Haemolysis
Test for Volatile oil
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Figure 1: Morphology of Cissus quadrangularis Linn.
Fig 2 : T.S. of Cissus quadrangularis Linn. Stem
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Figure 3: Powder analysis of Cissus quadrangularis Linn. Stem
End
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