MORPHO-ANATOMICAL AND PHYSICOCHEMICAL EVALUATION OF VITEX PINNATA LINN LEAVES

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Thenmozhi et al. European Journal of Biomedical and Pharmaceutical Sciences
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483
MORPHO-ANATOMICAL AND PHYSICOCHEMICAL EVALUATION OF VITEX
PINNATA LINN LEAVES
S. Thenmozhi1* and U. Subasini2
1*PRIST University, Vallam, Thanjavur – 613 403, Tamilnadu, India. / Dept of Pharmacognosy, Swamy Vivekanandha
College of Pharmacy, Elayampalayam, Tiruchengode, Namakkal (D.T), Tamilnadu, India.
2Dept of Pharmacognosy, Swamy Vivekanandha College of Pharmacy, Elayampalayam, Tiruchengode, Namakkal
(D.T), Tamilnadu, India.
Article Received on 19/02/2016 Article Revised on 09/03/2016 Article Accepted on 29/03/2016
INTRODUCTION
Vitex pinnata Linn. (Family: Verbenaceae) is a small to
medium-sized evergreen tree, up to 25-30 m tall[1,2],
often with a crooked bole, up to 70 cm in diameter at
breast height; bark surface smooth, shallowly fissured or
flaky, pale grey to yellowish brown, inner bark pale
yellow to bright orange; branches quadrangular, crown
often spreading. Leaves opposite, compound, 3-5
foliolate; leaflets and petioles pubescent below; lateral
leaflet sessile or nearly so, elliptic, 10-20 cm long. The
plant has been reported to have various ethanomedicinal
applications.[3-5] In Traditional medicine, the plant is
used to expel intestinal worms, as analgesic, anti-
inflammatory[6], antipyretic, wound healing[7], anti-
oxidant, antibacterial and stomach ache.[8] In Brunei, the
young leaf shoots are eaten raw to treat hypertension and
fever. A root-tea is taken for backache, body ache and
fatigue.[9] Leaves are used to cure fever and wounds
while the bark scrapings are applied to wounds and used
as charm for convulsions.[10] A bark extract is taken for
the treatment of jaundice.
MATERIALS AND METHODS
Plant material
The leaves of selected plant were collected in the month
of July 2013 from Tirunelveli, Tamilnadu and were
identified and authenticated by Retired Scientist
Dr.V.Chelladurai, M.Sc., Ph.D., Research Officer –
Botany, Central Council for Research in Ayurveda &
Siddha, Govt. of India. Tirunelveli, Tamil Nadu, [the
Voucher specimen No. SVCP/Ph.Cog/118], has been
deposited in the Department of Pharmacognosy, Swamy
Vivekanandha College of Pharmacy, Tiruchengode,
Namakkal (D.T), Tamil nadu.
Macroscopic evaluation
The organoleptic and macroscopic characters of Vitex
pinnata Linn leaves like colour, odour, shape, size, taste,
ect,. were evaluated.
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Research Article
ejbps, 2016, Volume 3, Issue 4, 483-492.
European Journal of Biomedical
AND Pharmaceutical sciences
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ISSN 2349-8870
Volume: 3
Issue: 4
483-492
Year: 2016
*Author for Correspondence: S. Thenmozhi
PRIST University, Vallam, Thanjavur – 613 403, Tamilnadu, India. / Dept of Pharmacognosy, Swamy Vivekanandha College of Pharmacy,
Elayampalayam, Tiruchengode, Namakkal (D.T), Tamilnadu, India.
ABSTRACT
Vitex pinnata Linn. (Syn: Vitex pubescens Vahl.) is a moderately sized tree of tropical Asia belonging to the
family Verbenaceae. Vitex pinnata is the tree with trifoliate leaves, gray to brown bark. The leaves are potential as
antioxidant; because of flavonoids, alkaloids & terpenoids content. In Traditional medicine, the plant is used to
expel intestinal worms, as analgesic, anti-inflammatory, antipyretic, wound healing, anti-oxidant, antibacterial and
stomach ache. In Brunei, the young leaf shoots are eaten raw to treat hypertension and fever. A root-tea is taken
for backache, body ache and fatigue. The present study deals with the morphological and microscopic evaluation
of the leaf and powdered leaf material and establishment of the quality parameters including physicochemical
evaluation. The microscopical studies revealed the presence of mesophyll tissue, peltate type, capitate type of
glandular trichomes, non- glandular trichomes, reticulate venation pattern, anomocytic stomata and ground tissue,
etc,. Physicochemical evaluations showed Moisture content 5.1 ± 0.04%w/w, Foreign organic matter 6.8 ±
0.36%w/w, Total Ash 10.33 ± 0.16%w/w, Acid insoluble ash 1.33 ± 0.15%w/w, Water soluble Ash 2.03 ±
0.25%w/w, Sulphated Ash 3.0 ± 0.06%w/w, Alcohol soluble extractives 19.6 ± 0.26%w/w, Water soluble
extractives 28.8 ± 0.15%w/w respectively. The preliminary phytochemical analysis revealed the presence of
carbohydrates, alkaloids, flavonoids, saponins, phenolic compounds, tannins, steroids, protein and amino acids.
The study which is helpful in the identification of authentic plant from other vitex species.
KEYWORDS: Vitex pinnata Linn., Macroscopical evaluation, Microscopical characters, Physicochemical
analysis, Phytochemical screening.

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Instruments used
Photographs of different magnifications were taken with
Nikon Labphot2 Microscopic Unit. For normal
observations bright field was used. For the study of
crystals, starch grains and lignified cells, polarized light
was employed. Since these structures have birefringent
property, under polarized light they appear bright against
dark background.[11]
Materials and methods for Anatomical studies
Collection of specimens
The plant specimens were collected from Tirunelveli,
Tamil Nadu, India. Care was taken to select healthy
plants and for normal organs. The required samples of
leaves were cut and removed from the plant and fixed in
FAA (Formalin-5ml + acetic acid-5ml + 70% Ethyl
alcohol-90ml). After 24hrs of fixing, the specimens were
dehydrated with graded series of tertiary –butyl alcohol
as per the schedule given by Sass, 1940.[12] The
specimens were castled into paraffin blocks.
Sectioning
The paraffin embedded specimens were sectioned with
the help of Rotary Microtome. The thickness of the
sections was 10-12µm. De-waxing of the sections was
done by customary procedure.[13] The sections were
stained with Toluidine blue as per the specific method.[14]
For studying the venation pattern and trichome
distribution, paradermal sections (sections taken parallel
to the surface of leaf) of leaf with 5% sodium hydroxide
or epidermal peeling by partial maceration employing
Jefferey`s maceration fluid was prepared. Glycerin
mounted temporary preparations were made for
macerated/cleared materials. Powdered materials of
different parts were cleared with NaOH and mounted in
glycerin medium after staining. Different cell component
were studied and measured.
Photomicrographs
Microscopic descriptions of tissues are supplemented
with micrographs wherever necessary. Photographs of
different magnifications were taken with Nikon Labhot 2
Microscopic Unit. For the study of crystals, starch grains
and lignified cells, polarized light were employed. Under
polarized light they appear bright against dark
background. Magnifications of the figures are indicated
by the scale –bars. Descriptive terms of the anatomical
features are as given in the standard anatomy books.[15,16]
Power microscopy
The dried leaves were powered and studied under
microscope. A pinch of powder was taken in a
microscopic slide, 1-2 drops of 0.1%w/v phloroglucinal
solution and a drop of concentrated hydrochloric acid
were added and covered with a cover slip. The
characteristics features of cell components were
observed and their photographs were taken using
photomicrography.
Physicochemical analysis
The dried powdered leaves was subjected to
physicochemical analysis including fluorescence
analysis[17], moisture content, total ash, water soluble ash,
acid insoluble ash, sulphated ash, alcohol soluble
extractive and water soluble extractive[18] to determine
the quality and purity of the plant materials.
Preparation of extracts
The leaves were dried under shade and then powdered
with a mechanical grinder. The powder was passed
through sieve no.40 and stored in air tight container for
further use. The dried powdered leaves of Vitex pinnata
Linn. is subjected to hydroalcoholic extraction in the
ratio of Ethanol: water as 70:30, adopting cold
percolation method. The extract is concentrated under
reduced pressure to yield semi-solid mass which is dried
in a desiccator and then subjected to preliminary
phytochemical analysis using different chemical
tests.[19,20 ]
RESULT AND DISCUSSION
Macroscopic study
Small to medium-sized evergreen tree up to 25-30m tall.
Pale grey to yellowish brown colour bark, smooth,
shallowly fissured or flaky, inner bark pale yellow to
bright orange colour. Branches quadrangular, crown
often spreading. Leaves opposite, compound, 3-5
foliolate; leaflets and petioles pubescent below; lateral
leaflet sessile or nearly so, elliptic, 10-20 cm long.
Flowers in terminal panicles, compact, pyramidal, blue
or lilac, aromatic stamens 4, 2 longer, 2 shorter. Ovary
superior, 2-4-chambered, with 1 filiform style having a
bifid stigma. Fruit a drupe, subglobose, 7-13 mm in
diameter, purplish- black when matured, sessile on the
often enlarged calyx, 1-4 seeded. Seed, obovoid or
oblong, lacking endosperm (Fig.1 & Fig. 2).
Fig. 1: Leaf & stem of Vitex pinnata Linn.

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Fig. 2: Tree of Vitex pinnata.
Microscopical Features of Vitex pinnata Linn leaves
i. Anatomy of the Leaf
The leaf consists of thick and prominent midrib and their
lamina. The midrib and lamina are studded with dense
epidermal trichomes. The midrib consists of thick,
conical adaxial part and wide semi circular abaxial part.
The midrib is 1.3 mm thick, the adaxial core is 300µm
wide and the abaxial part is 1.35mm wide. The epidermal
layer of the midrib consists of small, circular thick
walled cells bearing both glandular and non-glandular
trichomes. The adaxial core includes homogenous,
circular thick walled parenchyma cells. The abaxial part
consists of 4 or 5 layers of collenchymatous cells in the
outer part and thin walled circular parenchymatous zone
in the inner part.
The midrib exhibits complex vascular system. It includes
a wide and deep bowl shaped main vascular strand and
the medullary group of isolated 7 or 8 circular vascular
strands. The entire system is surrounded by fairly thick
cylinder of thick walled lignified fibres (Fig. 3.2). The
main bowl shaped vascular strand consists of numerous
short or long parallel lines of xylem elements alternating
with narrow passages of fibres. The xylem elements are
angular, wide and thick walled. Along the lower
boundary of xylem arc occur several masses of phloem
elements. The medullary vascular strands are circular
and collateral. Each strand includes an arc of wide thick
walled angular xylem elements and a large mass of
phloem elements.
ii. Lamina
The lamina is bilaterally symmetrically measuring
140µm thick. The adaxial epidermis includes cylindrical
fairly wide thin walled cells with thick cuticle. The
abaxial epidermis is narrow and the cells are square
shaped or cylindrical. The mesophyll tissue includes an
adaxial zone of 3 horizontal rows of cylindrical palisade
cells. The palisade cells are compact and densely
chlorophyllous. The abaxial part of the lamina includes 7
or 8 lobed somewhat compact spongy parenchyma cells.
There are vertical pillars of lateral veins which include
spindle shaped mass of thick walled xylem elements with
parenchymatous bundle sheath. The bundle sheath
extends both adaxially and abaxially up to epidermal
layers (Fig. 4.1).
Fig. 3.1: T.S of leaf through midrib.
Fig. 3.2: T.S of midrib enlarged.
(AbS- Abaxial strand, AdC- adaxial cone, AdS- Adaxial
strand, Col- Collenchyma, Ep- Epidermis, ETr-
Epidermal Trichomes, GT- Ground tissue, La- Lamina,
Pa-Parenchyma, Ph- Phloem, SC- Sclerenchyma, X-
Xylem).
Fig. 4.1: T.S of lamina.

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Fig. 4.2: T.S of leaf margin.
(AbE- Abaxial Epidermis, AdE- Adaxial epidermis, Cu-
Cuticle, GTr- Glandular Trichome, LM- Leaf margin,
LV- Lateral vein, MT- Mesophyll tissue, PM- Palisade
Mesophyll, SC- Secretory Canal, SM - Spongy
Mesophyll).
iii. Leaf margin
The marginal part of the lamina is slightly curved down
and measures 120µm thick. The epidermal layer of the
leaf margin includes small much thick walled cells. The
inner portion of the marginal part includes a compact
mass of sclerenchymatous cells. The remaining portion
exhibits normal differentiation of mesophyll tissues.
Glandular and non-glandular trichomes are also in leaf
margin (Fig. 4.2).
iv. Epidermal trichomes
Three types of epidermal trichomes are seen on the
lamina.
a. Peltate type of glandular trichomes
It is wide, bowl shaped gland attached on the epidermal
layer in a concavity. The gland consists of basal cells and
a wide bowl shaped peltate head. At the bottom of the
peltate body occur a group of secretory cells which stain
darker (Fig. 5.1). On maturity the secretary cells break
and fuse forming a wide elliptical secretory liquid at the
bottom of the head (Fig. 5.2). The peltate trichomes are
20µm height and 70µm wide.
Fig. 5.1: T.S of lamina bearing peltate type of
glandular trichome.
Fig. 5.2: Peltate trichome enlarged.
(AdEp- Adaxial Epidermis, BC- Basal Cell, Ep-
Epidermis, PH- Peltate Head, SC-Stalk Cell, SL-
Secretory Liquid, PM- Palisade Mesophyll).
b. Capitate type of glandular trichomes
The capitates trichomes are short with one celled stalk
and spherical darkly stained terminal part. These
trichomes may occur either on the adaxial epidermis
(Fig. 6.2) or on the abaxial epidermis (Fig. 6.3).The
gland is 45µm in height and 25µm in thickness.
c. Non- glandular trichomes
The non-glandular trichomes are more abundant and they
are multicellular, uniseriate and unbranched. The
trichome is broader at the base and gradually tapers
towards the tip into curved pointed apex. The trichome
has very thick lignified walls and narrow lumen. The
trichome is about 100µm in height and 25µm thick (Fig.
6.1).
Fig. 6.1: Non glandular trichome on the lamina.
Fig. 6.2 Adaxial glandular trichome.

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Fig. 6.3: Abaxial glandular trichome.
(AbE- Abaxial Epidermis, AbGT- Abaxial Glandular
Trichome, AdE- Adaxial Epidermis, AdGT- Adaxial
Glandular Trichome, NGT- Non-Glandular Trichome,
PM-Palisade Mesophyll, SP- Spongy Parenchyma).
v. Venation pattern
In the cleared preparation of the lamina, the surface view
of venation system was studied. The venation is densely
reticulate. The veins include thick secondary veins, thin
tertiary veins which form rectangular or square shaped
vein islets. The boundary of the vein islets includes thick
and straight veins. The vein terminations are short, either
unbranched and straight or branched ones (Fig. 7.2 &
7.3). Sometimes the vein terminations are long and
curved into hook (Fig. 7.2).
Fig. 7.1, 7.2 &7.3: Venation system of the lamina
showing vein islets and vein termination. (VI-Vein
Islet, VT-Vein Termination).
vi. Petiole
Both distal and proximal regions of the petiole where
studied. The distal part of the petiole is wide and
semicircular with flat adaxial part and short, thick,
conical lateral wings (Fig. 8.1).The proximal petiole is
also semicircular but the adaxial side is slightly raised
and wings are short and much wide (Fig. 8.2).
Fig. 8.1: T.S. of distal part of the petiole. Fig. 8.2: T.S.
of the proximal part of the petiole. (AdS-Adaxial Side,
GT-Ground Tissue, MB-Median Bundle, VC-Vascular
Cylinder, W-Wing, WB-Wing Bundle).
The distal petiole is 1.6mm thick and 2.4mm wide. The
proximal petiole is 1.75mm thick and 2.2mm wide.
The petiole has thin layer of epidermis and the epidermal
cells are small and thick walled. The ground tissue of the
distal part includes outer zone of small thick walled
circular cells and inner zone of fairly large thin walled
parenchyma cells. The vascular system is complex one
comprising outer main system and the central medullary
system. The outer system forms wide omega shaped
outline and it includes several radial files of xylem
elements all along the outline of the strand. The xylem
elements are elliptical, thin walled with wide lumen.
Mixed with the xylem elements are xylem fibres. The
outer boundary of xylem strand includes fairly thick
continuous layer of primary phloem and the vascular
cylinder is surrounded by thick sclerenchymatous sheath.
The central medullary bundles are numerous and each
bundle is circular with central core of xylem elements
surrounded by phloem elements.
The distal part of the petiole has prominent conical wing
and large semicircular, collateral vascular bundles (Fig.
9.1). The wing bundle has 4 short lines of xylem
elements and an arc of phloem elements. The entire
bundle is surrounded by a thick sclerenchymatous bundle
sheath. The central medullary bundles are diffuse in
distribution and they are large and collateral. Each
bundle consists of 3 to 5 short parallel lines of xylem
elements and wide mass of phloem elements. The ground
tissue in the central part has dense accumulation of
tannins (Fig. 9.2). The ground tissue of the proximal
peptide consists of outer thick cylinder of
sclerenchymatous elements and inner ground tissue of

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thin walled parenchymatous cells. The main bowl shaped
vascular strand consists of numerous, long, narrow lines
of xylem elements and discontinuous masses of phloem
elements located along the outer periphery. There is no
sclerenchymatous cylinder encircling the entire vascular
system. The medullary bundles are diffuse in distribution
and circular in outline. These bundles have a few xylem
elements and a wide mass of phloem elements and a
wide mass of phloem elements. Xylem and phloem are
collateral in arrangement. The ground tissue in the
medullary region contains dense accumulation of tannin.
Fig. 9.1: Adaxial part of the distal petiole showing
wing and wing Bundle.
Fig. 9.2: Medullary bundles of the distal petiole.
(AdS- Adaxial Side, Ph- Phloem, Sc- Sclerenchyma, W-
Wing, X- Xylem).
Powder microscopy
Following inclusions are seen in the powder preparation.
Epidermal peeling of the lamina is often seen in the
powder (Fig. 10.1 & 10.2). The peeling displays the
surface features of the epidermal cells and stomatal
morphology.
Fig. 10.1: Epidermal peeling of the lamina as seen in
surface view showing epidermal cells and stomata.
(SC- subsidiary cells, St- stomata).
Fig. 10.2: Four stomata and epidermal cells enlarged.
The epidermal cells are small, polyhedral and thick
walled. The anticlinal walls are slightly wavy (Fig. 10.1).
The stomata are densely distributed. They are random in
orientation. The stomata are anamocytic type. The guard
cells are surrounded by ordinary epidermal cells. The
guard cells are circular in outline and measure 12×15µm
in size.
Epidermal trichomes
Epidermal trichomes are fairly abundant in the powder.
There are two types of trichome seen in the powder.
a) Non –glandular trichomes (Fig. 11.1, 11.2 & 11.3)
The non glandular trichomes are multi cellular,
unbranched and uniseriate. The trichomes may be two
to five celled. The cells are vertically elongated and thick
walled. The cell-lumen is wide and possess some
granular inclusions. The trichomes are gradually tapering
into pointed tip. The longest trichome is 210µm long
and 20 µm thick.
Fig. 11.1: Non glandular trichomes attached on the
vein.

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Fig. 11.2: Non glandular trichomes – 2 celled and 3
celled.
Fig. 11.3: 5 celled curved trichomes. (NGTr – Non
Glandular Trichome).
b) Glandular trichomes (Fig. 12.1 & 12.2)
The glandular trichomes are capitate type. They have
short stalk and large spherical or obconical body (Fig.
12.1). In surface view, the body part of the trichome is
circular with central part representing the stalk of the
gland (Fig. 12.2). The body of the trichome is 80µm in
diameter.
Fig.12.1: Detached glandular trichomes as seen in
lateral view and surface view.
Fig.12.2: A glandular trichome as seen in surface
view showing central stalk cell and circular body
cell. (BC – Body Cell, GTr – Glandular Trichome, Sc –
Stalk cell).
Fibre (Fig. 12.3)
Thick and long fibers are less frequently seen in the
powder. The fibre has thick lignified walls, wide lumen
and tapering ends. Some inclusions are seen within the
lumen of the fibre.
Fig. 12.3: A detached fibre showing wall thickness
and cell lumen. (Fi – fibre).
Physicochemical analysis of crude drug
Physicochemical evaluations showed Moisture content
5.1 ± 0.04%w/w, Foreign organic matter 6.8 ±
0.36%w/w, Total Ash 10.33 ± 0.16%w/w, Acid insoluble
ash 1.33 ± 0.15%w/w, Water soluble Ash 2.03 ±
0.25%w/w, Sulphated Ash 3.0 ± 0.06%w/w, Alcohol
soluble extractives 19.6 ± 0.26%w/w, Water soluble
extractives 28.8 ± 0.15%w/w respectively (Table 1).
Table 1: Physicochemical analysis of Vitex pinnata
Linn leaves.
S.No.
Parameters
Value obtained on dry
weight basis (%w/w)
1
Moisture content
5.1 ± 0.04
2
Foreign organic
matter
6.8 ± 0.36
3
Total Ash value
10.33 ± 0.16
4
Acid insoluble ash
1.33 ± 0.15
5
Water soluble Ash
2.03 ± 0.25
6
Sulphated Ash
3.0 ± 0.06
7
Alcohol soluble
extractive
19.6 ± 0.26
8
Water soluble
extractive
28.8 ± 0.15
Fluorescence analysis
The powder drug with different chemical reagents
showed different colour when seen on naked eye. The
different colour observed shows the presence of different
type of phytoconstituents (Table 2). Many drugs
fluorescence when their powder is exposed to ultraviolet
radiation. It is important to observe all materials on
reaction with different chemical reagents under U.V.
light. The fluorescence characteristics of powdered drug
were studied under U.V. light after treating with different
chemical reagents are reported. (Table 3).
Table 2: Reaction of powdered drug with different
reagents.
Treatment
Colour
Powder as such
Pale green
Powder + conc. H2SO4
Brownish black
Powder + conc. HNO3
orange colour
Powder + conc. HCL
Yellowish brown
Powder + 5% Iodine
Reddish brown
Powder + 5M NaOH
Brownish yellow
Powder + glacial acetic acid
Yellowish green
Powder + 80% H2SO4
Light yellow

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Table 3: Fluorescence Analysis of powdered drug under UV light.
Preliminary phytochemical screening
The percentage yield of hydroalcoholic extract of Vitex
pinnata Linn leaves was found to be 28.46 %w/w with
semi-solid masses in dark brown colour. The
hydroalcoholic extract were subjected to phytochemical
screening which reveals the presence of various
pharmacological active compounds such as
carbohydrates, alkaloids, phenolic compounds, tannins,
proteins and amino acids, saponins, steroids and
triterpeniods, flavonoids shown in Table 4. Thin layer
chromatography was carried out with different mobile
phases and the best solvent system was chloroform and
ethyl acetate (6:4) and revealed 6 spots with different Rf
values under UV light at 366nm (Table 5).
Table 4: Preliminary Phytochemical screening of Vitex pinnataLinn. Leaves.
Reagents
Day Light
UV Light
Drug powder
Pale green
Dark Green
Drug Powder + 1M NaOH
Orange brown
Greenish yellow
Drug Powder + alc. 1M NaOH
Pale green
Pale green
Drug powder + 1M HCl
Pale green
Green
Drug powder + 50% HNO3
Yellowish orange
Yellowish green
Drug powder + 5% Fecl3
Yellowish brown
Dark green
Drug powder + 80% H2SO4
Yellowish brown
Greenish yellow
Drug powder + water
Pale green
Pale green
Drug powder + conc. H2SO4
Crimson red
Black colour
Drug powder +Chloroform
Pale green
Pale green
Drug powder +Benzene
Pale green
Pale green
Drug powder +Acetone
Pale green
Pale green
Drug powder +Alcohol
Pale green
Pale green
S.No.
PLANT CONSTITUENTS
IDENTIFICATION TESTS
HYDRO
ALCOHOLIC EXTRACT
1.
Alkaloids
Mayer’s test
-
Hager’s test
+
Wagner’s test
+
Dragendroff’s test
-
2.
Carbohydrates
Molisch’s test
+
Fehling’s test
+
Benedict’s test
+
Barfoed’s test
-
Selwinoff’s test
-
3.
Glycosides
Borntrager’s test
-
Legal’s test
-
Keller killiani test
-
Conc.H2So4
-
4.
Phenolic compounds &
Tannins
Ferric chloride test
+
Lead acetate test
+
Ellagic acid test
-
5.
Protein and amino acids
Millon’s test
+
Ninhydrin test
-
Biuret test
+
Xanthoprotein test
-
6.
Saponins
Foam test
+
7.
Steroids and triterpenoids
Libermann burchard test
+
Salkowski test
+
8.
Fixed oils and Fats
Spot test
-
Saponification test
-
9.
Flavonoids
Shinoda test
+
Alkaline reagent test
+
Conc.H2so4
+
Ferric chloride test
+
Fluorescence test
+
10.
Mucilage and gums
With 90% alcohol
-

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Table 5: TLC of hydroalcoholic extract of Vitex pinnata Linn.
S.No
Name of the
Extract
Solvent system
Spots
observed
Rf values
1.
Hydroalcoholic extract
Chloroform : Ethyl acetate :
Formic acid : Water
(9:0.3:0.2:1)
4
0.50
0.60
0.73
0.95
2.
Chloroform : Toluene : Ethyl
acetate (6:3:1)
5
0.09
0.10
0.41
0.71
0.79
3.
Toluene : Ethyl acetate : Formic
acid (9:0.5:0.5)
4
0.05
0.07
0.17
0.24
4.
Chloroform : Ethyl acetate
(6:4)
6
0.07
0.16
0.30
0.96
0.94
0.98
CONCLUSION
The present study is used to investigate the
pharmacognostical, physical constants and preliminary
phytochemical screening of Vitex pinnata Linn. leaves
provided useful information about its correct identity and
evaluation. The macroscopical studies revealed the
morphological character of different parts of the plant.
The microscopical studies revealed the presence of
mesophyll tissue, peltate type, capitate type of glandular
trichomes, non- glandular trichomes, reticulate venation
pattern, anomocytic stomata and ground tissue, etc,.
Various physicochemical parameters such as ash values,
extractive values, foreign organic matter, moisture
content and fluorescence analysis were determined.
Phytochemical screening is also useful to isolate the
pharmacologically active principles present in the drug.
The other parameters observed are also useful for the
future identification of the plant and serves as a standard
monograph for identification and evaluation of plant.
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