Phytochemical screening of Hemigraphis colorata (Blume) H.G. Hallier

Abstract

Hemigraphis colorata (Blume) H.G. Hallier (syn: Hemigraphis alternata, family: Acanthaceae), is an exotic plant adapted to India. It is a versatile low-creeping perennial herb mostly valued as an ornamental plant. Since remote past, the leaves are ground into a paste and applied on fresh cut wounds to promote wound healing. Clinical studies also highlight its significant antimicrobial, anti-diabetic and anti oxidant activities. The present investigation present study was focused to evaluate the pharmacognostic and preliminary phytochemical properties of H. colorata leaves. The morphological and anatomical characters, quantitative microscopy, powder microscopy and the behavior of powdered sample with different reagents were recorded. Pharmacognostic study of crude drug would be essential for any advanced pharmaceutical research on this plant.

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Journal of Pharmacognosy and Phytochemistry 2021; 10(6): 360-363
E-ISSN: 2278-4136
P-ISSN: 2349-8234
www.phytojournal.com
JPP 2021; 10(6): 360-363
Received: 16-09-2021
Accepted: 18-10-2021
Devi Priya M
Department of Botany,
St. Thomas College, Ranni,
Pazhavangadi PO,
Pathanamthitta, Kerala, India
Vinod Kumar TG
Department of Botany,
St. Thomas College, Ranni,
Pazhavangadi PO,
Pathanamthitta, Kerala, India
Francis Mathew
Department of Botany,
St. Thomas College, Ranni,
Pazhavangadi PO,
Pathanamthitta, Kerala, India
Corresponding Author:
Devi Priya M
Department of Botany,
St. Thomas College, Ranni,
Pazhavangadi PO,
Pathanamthitta, Kerala, India
Phytochemical screening of Hemigraphis colorata
(Blume) H.G. Hallier
Devi Priya M, Vinod Kumar TG and Francis Mathew
Abstract
Hemigraphis colorata (Blume) H.G. Hallier (syn: Hemigraphis alternata, family: Acanthaceae), is an
exotic plant adapted to India. It is a versatile low-creeping perennial herb mostly valued as an ornamental
plant. Since remote past, the leaves are ground into a paste and applied on fresh cut wounds to promote
wound healing. Clinical studies also highlight its significant antimicrobial, anti-diabetic and anti oxidant
activities. The present investigation present study was focused to evaluate the pharmacognostic and
preliminary phytochemical properties of H. colorata leaves. The morphological and anatomical
characters, quantitative microscopy, powder microscopy and the behavior of powdered sample with
different reagents were recorded. Pharmacognostic study of crude drug would be essential for any
advanced pharmaceutical research on this plant.
Keywords: Hemigraphis colorata, Leaf, Phytochemical
Introduction
Hemigraphis colorata is an excellent indoor and outdoor plant, chiefly grown because of its
attractive and vivid foliage. It prostrates and spreads with rooting stems when grown on
ground, and on hanging baskets it cascades over beautifully. It is also used to decorate
aquariums and goldfish bowls. The plant is well adapted to live in tropical climate. The plant
is known by several vernacular names such as Aluminium plant, Cemetary plant, Metal leaf,
Red flame Ivy, Waffle plant, Java Ivy etc. In Kerala it is known as ‘murikootti’ or ‘murian
pacha’ (Fig 1).
Hemigraphis is a versatile low-creeping perennial herb that reaches a height of 15 to 30 cm.
The leaf has metallic purple lustre on upper surface and a solid dark purple on ventral side.
The leaves are opposite, ovate to cordate, serrate-crenate, about 2 to 8 cm long and 4 to 6 cm
wide, bearing well-defined veins. It blooms irregularly throughout the year in the tropics.
Flowers are small (1 to 1.5 cm diameter), five lobed, bell shaped with imbricate bracts. These
are white in colour with faint purple marks within and appear in terminal 2 to 10 cm long
spikes. Capsules are small, slender, oval, linear and light green in colour. Seeds are small, flat
and white in colour [1-3].
Fig 1: Habit of H. colorata

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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Pharmacology and Phytochemistry
Hemigraphis is a therapeutic plant with immense power to
cure vitiated pitta, fresh wound, cuts, ulcers, inflammations
and skin complaints. Traditionally, the leaves are consumed to
mend gall stones and excessive menstruation. In Vanuatu, sap
of leaf buds are squeezed in water and drunk at dawn for 4
days as contraceptive and to induce sterility [4]. In Java, leaves
are used to treat bloody dysentery and piles. It is also has
diuretic ability. In folk medicine, the leaf juice is applied
directly on open wound to stop bleeding [5, 6] and is used
internally to cure anemia.
Phytochemicals are variety of secondary metabolites which
provide the curative property. The phenolic compounds in the
benzene extract of H. colorata leaves have showed its activity
against Acinetobacter species and Streptococcus aureus [7].
The phenolic acids such as chlorogenate, cinnamate,
coumarate, gallate and ferulate present in the plant acts as
pro-oxidants and exhibits free radical scavenging activity [8].
The steroids and coumarins present in the extract provide
anti-diabetes activity [9]. The crude leaf paste promotes
excision wound healing [10, 11]. In mice, the leaf paste provides
faster wound contraction and epithelialisation but oral
administration is seen ineffective [12]. The excision and
incision wound model studies revealed that methanolic extract
is comparable to standard reference Vokadine [13]. The herbal
scaffold made from chitosan was highly haemostatic and can
be effectively applied for infectious wounds [14].
Materials and Methods
The plants were collected from the natural habitat, Erumeli,
Kottayam, Kerala and was positively identified and confirmed
by the herbarium in the Department of Botany, University of
Kerala.
Pharmacognostic Studies
Morphological studies were performed by physical
evaluation. For micro-characterization, free hand sections of
about 10-20 μm thickness of leaves were made and stained
with safranine (0.5%). The micro slides preparation was
further observed in an image analyzer (Olympus BX 51) and
the anatomical peculiarities were photo documented.
As a part of quantitative microscopy, stomatal number,
stomatal index and vein islet number were determined by
using fresh leaves of the plant (Wallis 1985) [15]. Stomatal
type and various leaf constants such as stomatal number,
stomatal index, vein islet number, and palisade ratio [16]
(Chase and Pratt 1949) were determined by using fresh
leaves. Organoleptic character like total ash, acid insoluble
ash and watersoluble ash values were also determined
(Kokashi et al., 1958; Daniel, 1991; Harborne, 1998) [17-19].
Behaviour of powder with different reagents was also noticed.
Preliminary Phytochemical Studies
For preliminary phytochemical studies extraction of crude
drug with different solvents was carried out by direct
extraction by cold maceration method using solvents viz.
acetone, methanol and distilled water as per increasing order
of their polarity. Presence of various phytoconstituents viz.,
alkaloids (Dragendorff’s test, Mayer’s test, Wagner’s test),
aminoacids (Ninhydrin test), anthocyanin, anthraquinones
(Borntrager’s test, Modified Borntrager’s test), carbohydrates
(Benedict’s reagent test, Fehling solution test, Molisch's test),
coumarins (Sodium hydroxide test), flavonoids (Shinoda test,
Ammonia test, Sodium Hydroxide test), flavones, glycosides
(Anthrone test, Benedict’s reagent test, Fehling solution test),
phenols (Ferric chloride test, Phosphomolybdic acid test),
phlobatannins (Hydrochloric acid test), protein (Biurete test,
Millon's test, Xanthoprotein test), quinines (Sodium
hydroxide test),quinines (Sulphuric acid test), reducing sugar
(Benedict’s test, Fehling test), resins (Turbidity test), saponins
(Foam test), steroids and terpenoids (Lieberman burchard test,
Salkowski test, Hirshonn reaction) and tannins (Braemer’s
test, Bromine water test) were tested.
Result and Discussion
The plants are playing an important role in the development
of novel drugs and well defined pharmacognostic parameters
and standards must be established before the inclusion of any
crude drug in herbal pharmacopoeia. Physical evaluation is
the major step employed in the identification and
standardization of crude drugs. It helps in the determination of
adulterants and confirms the genuineness of crude drug.
Table 1: Quantitative microscopic examination of Hemigraphis
colorata leaf
Determinations
Value
Palisade ratio
2.75
Stomatal Index (Upper epidermis)
8.65
Stomatal Index (Lower epidermis)
22.34
Vein islet number
2.3
Fig 2: Stomatal type of H. colorata
The leaves have metallic purple coloured dorsal surface and a
solid purple colour on ventral surface. They are opposite,
ovate to heart-shaped, with scalloped edges. The leaves show
an area ranges between 17.1- 22.5cm2, length 5.1-7.6 cm and
width vary from 3.9 cm to 4.8 cm. On upper surface of the
leaves trichomes are present. Leaf peeling from upper and
lower side showed diacytic stomata, in which cross walls are
transverse to the long axis of the stoma (Fig- 2). The stomatal
index on upper epidermis is 8.65 and lower is 22.34. The
palisade ratio is calculated as 2.75. The vein islet number and
vein termination number are 2.3 respectively (Table 1). Vein
islet can be used for proper identification of plants and
standardization of crude drug to prevent from adulteration of
drug in powdered form.
Table 2: Physico-chemical characteristics
Ash value
Leaf
Total Ash (%)
11.8
Acid insoluble ash (%)
1.18
Water insoluble ash (%)
10.1
Water soluble ash (%)
2.22

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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Table 3: Organoleptic characters
Character
Leaf Powder
Colour
Ash green
Odour
Mouldy
Size
Coarse
Taste
Bitter
Texture
Coarse
C.S. of leaf showed single layered epidermis covered with
cuticle. The upper region of the leaf surface also contains
multicellular non-glandular trichomes. Mesophyll is
differentiated into upper radially elongated and compactly
packed single layered palisade tissue and loosely arranged
spongy tissue. In between palisade and spongy tissues many
layered compactly packed thin walled parenchymatous cells
(Fig-3). Well define bundle sheath cells without chloroplast is
seen surrounding vascular tissue. Xylem is composed of
trachieds, vessels, fibres and parenchyma. Phloem is present
along with xylem and is composed of non-lignified cells.
Polygonal cells of chlorenchyma present below upper
epidermis and above lower epidermis.
C.S. of leaf showed single layered epidermis covered with
cuticle. The upper region of the leaf surface also contains
multicellular non-glandular trichomes. Mesophyll is
differentiated into upper radially elongated and compactly
packed single layered palisade tissue and loosely arranged
spongy tissue. In between palisade and spongy tissues many
layered compactly packed thin walled parenchymatous cells
(Fig-3). Well define bundle sheath cells without chloroplast is
seen surrounding vascular tissue. Xylem is composed of
trachieds, vessels, fibres and parenchyma. Phloem is present
along with xylem and is composed of non-lignified cells.
Polygonal cells of chlorenchyma present below upper
epidermis and above lower epidermis.
As the part of physic-chemical studies, ash value
determination was carried out and complied (Table 2). The
official ash values are of key requisite in determination of the
purity of powdered drugs. Organoleptic characters refer to
evaluation of drugs with the use of sense organs and were
recorded (Table 3). Detection of colour variation of the
powdered samples under day light is also a way to identify its
purity (Table 4).
Table 4: Behaviour of the H. colorata powdered leaf drug with
various reagents
Reagents
Colour formation
Powder+ Distilled Water
Bluish grey
Powder+ 5% FeCl3
Lemon yellow
Powder+ l% Acetic acid
Magenta Pink
Powder+ 5% KOH
Royal Ivory
Powder+ 5% NaOH
Solemn yellow
Powder+ Conc. HCl
Rouge red
Powder+ Conc. H2SO4
Reddish brown
Powder+ Conc. HNO3
Happy celebration
Powder+ N/10 Iodine soln.
Yellowish red
Powder+ Ammonia soln.
Light reddish green
The pharmacological action of the crude drug is largely
depends on the metabolites present in it. The qualitative
screening by using various extracts revealed the presence of a
wide range of phyto-constituents (Table 5). The test was
positive to alkaloids, carbohydrates, flavonoids, phenols,
proteins, resins steroids and terpenoids. This is in conformity
with earlier work [20, 21].
Conclusion
The world is now focused more towards the utilization of
plants and plant related compounds to safe guard human
health, which is attributed mainly by their phyto-compounds.
The present study is a preliminary attempt to provide detailed
information on the biological, biochemical and physical
properties of leaves of Hemigraphis colorata. The
quantitative determination of pharmacognostic parameters can
be used to differentiate the H. colorata from closely related
species and also to become aware of adulteration.
Table 5: Phtochemical characterization of crude drug of H. colorata leaf powder
Phytochemical
Name of Test
Acetone
Methanol
Water
Alkaloids
Dragendorff’s test
Present
Present
Present
Mayer’s test
Present
Present
Present
Wagner’s test
Present
Present
Present
Aminoacid
Ninhydrin test
Absent
Absent
Absent
Anthocyanin
Sodium hydroxide test
Absent
Absent
Absent
Sulphuric acid test
Absent
Absent
Absent
Anthraquinones
Borntrager’s test
Absent
Absent
Absent
Modified Borntrager’s test
Absent
Absent
Absent
Carbohydrate
Bendict’s reagent test
Present
Present
Present
Fehling solution test
Present
Present
Present
Molisch’s test
Present
Present
Present
Coumarins
Sodium hydroxide test
Absent
Absent
Absent
Flavonoids
Ammonia test
Absent
Present
Present
Lead acetate test
Absent
Absent
Present
Shinoda test
Absent
Absent
Present
Flavones
Sodium hydroxide test
Absent
Absent
Present
Sulphuric acid test
Absent
Absent
Present
Glycosides
Anthrone test
Absent
Absent
Absent
Bendict’s reagent test
Absent
Absent
Absent
Fehling’s test
Present
Present
Present
Phenols
Ferric chloride test
Present
Present
Present
Phosphomolybdic acid test
Present
Present
Present
Phlobatannins
Hydrochloric acid test
Absent
Present
Present
Proteins
Biuret test
Absent
Absent
Present
Million’s test
Absent
Absent
Present
Xanthoprotein test
Absent
Absent
Present

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Journal of Pharmacognosy and Phytochemistry http://www.phytojournal.com
Quinine
Sodium hydroxide test
Present
Present
Present
Quinone
Suphuric acid test
Present
Absent
Absent
Resin
Turbidity test
Absent
Absent
Present
Saponins
Foam test
Absent
Absent
Absent
Steroids/ Terpenoids
Lieberman Burchard test
Absent
Absent
Present
Salkowski test
Absent
Present
Present
Hirshonn reaction
Absent
Absent
Absent
Tannins
Braemer’s test
Absent
Absent
Absent
Bromine water test
Absent
Absent
Absent
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