Pharmacognostical
analysis of Fruit of Toddalia asiatica
(L.) Lam. (Rutaceae)
M. Arul, N Sami Veerappa*
Department of
Botany, Periyar EVR College (Autonomous), Tiruchirappalii - 620 023, Tamil Nadu, India.
*Corresponding Author E-mail: prof.nsveerappa@gmail.com
ABSTRACT:
Toddalia asiatica is a reputed ever green tree
belonging to the family Rutaceae; commonly known as
Indian tulip tree. The plant is distributed tropical regions and coastal forest
in India. It is well known and all the parts are used in Indian system of medicine.
The plant has been used as malaria, toothaches and bronchial pains. The scientific
parameter is necessary to identify the exact plant material and to find its
quality and purity. The present study deals with various pharmacognostical
examinations like organoleptic or macroscopical
characters, microscopical or anatomical studies were
carried out and the parameters were reported. These studies indicated the
possible information for correct identification and standardization of this
plant material.
KEYWORDS: Toddalia asiatica, fruit, microscopy
and pahrmacognostical evaluation.
INTRODUCTION:
Toddalia asiatica (L.) Lam. Is known as ‘Milagarani’ in Siddha system of medicine it is knowm as Kanchana in Ayurveda. Toddalia asiatica a
very variable rambling, prickely, sarmentose
shrub, distributed almost throughout India, ascending to an altitude of 2,
500m. In south India, the plants are very common in the Nilgiri
and palani hills and also in the scrubby jungles of
Orissa. In the plains, particularly in dry situations, the plant assumes the
form of a low shrub with smaller and narrower leaflets. Plant contains coumarins toddaalone, toddanol norbraylin, and 5, 7,
8-trimethoxy coumarin. Toddalia is a monotypic
genus of family Rutaceae1, 2. It is a flowering plant in the citrus
family containing the single species Toddalia
asiatica commonly known as orange climber2.
The
botanical synonyms of Toddalia asiatica (L.) Lam. includes Paullinia asiatica,
Toddalia aculeate, Toddalia
effusa and Toddalia
ambigua. It is a liana with woody, corky, thorny
stems that climb on trees, rea- ching
up to 10 meters in length. It has yellow-green flowers, shiny green
citrus-scented leaves, and orange fruits about half a centimeter in diameter
that tastes like orange peel. The seeds are dispersed by birds and monkeys that
eat the fruits within the forests3.
Toddalia asiatica, is native to many
countries in Africa and Asia and it grows in forested riparian habitat with
high rainfall4. It is used as a folklore remedy for various ailments
in many areas of the world. T. asiatica
is used in treatment of several diseases that includes malaria5, 2 stomach
ache, chest pains, sore throat, food poisoning2, cough, indigestion,
nasal and bronchial pains6. Extracts of the plant have demonstrated
antiviral activity against H1N1 influenza in the laboratory7. The
harvest of this slow-growing plant from the wild for medicinal use may cause
its populations to decline and protocols for domestication or propagation of
the tree are being researched4.
In
Africa the plant is known by various local names: Ajua,
Nyalwet (Luo), Katemwe (Tugen), Mkuro, Chikombe za Chui (Digo), Mdakakomba (Swahili), Mwikunya
(Kikuyu), Olebarm-onyo (Maasai),
Gwambadzi (Vhavenda SA), Ranklemoentjie (Afrikaan). The
mode of propa-gation is by seed or root suckers4.
Remedy for various ailments in many areas of the
world. T. asiatica is used in treatment of several diseases that includes
malaria2, 5, stomach ache, chest pains, sore throat, food poisoning2, cough,
indigestion, nasal and Extracts of the plant have demonstrated antiviral activity against H1N1 influenza in the
laboratory7. The harvest of this slow-growing
plant from the wild for medicinal use may cause its populations to decline and protocols for domestication
or propagation of the tree are being researched4.
Medicinal plants are
well-known natural sources for the treatment of various diseases since
antiquity. About 20, 000 plant species used for medicinal purposes are reported
by World Health Organisation8. Despite emphasis being put in
research of synthetic drugs, a certain interest in medicinal plants has been
reborn, in part due to the fact that a lot of synthetic drugs are potentially
toxic and are not free of side effects on the host. This has urged
microbiologists all over the world for formulation of new antimicrobial agents
and evaluation of the efficacy of natural plant products as a substitute for
chemical antimicrobial agents9.
Toddalia asiatica Lam. (Family - Rutaceae) is
a straggler, known as “Milakarani” in Tamil, Kanchana‟ in Sanskrit, and „Kanj‟
in Hindi, used in Indian systems of medicine. There are two varieties; Toddalia asiatica var.
gracilis and var. floribunda and they are found in
the west and southern parts of India. Plants of var. gracilis
are generally found in denuded slopes, waste lands and lower hills, whereas
that of var. floribunda are found only on high altitudes. This plant grows well
in clay soils and is widespread in Africa, Asia and Madagascar10. Milakarani has gained popularity amongst traditional health
practitioners for treating numerous ailments. Amongst the documented ethnomedicinal uses, the fruit of this plant is known to
have been popularly applied in treating malaria, particularly in East Africa2.
The plant has been used in the curing of diseases like paralysis, malarial and
intermittent fever, dyspepsia, colic, diarrhea, cough, bronchitis, nausea,
wounds, contaminated ulcers, epilepsy, gonorrhoea and
general debility12, 13. These therapeutic values are due to the
presence of coumarins, quinoline
and benzophenanthridine alkaloids14.
Several compounds were isolated from various parts and different extracts of T.
asiatica. In 1965, Pakrashi
and Bhattacharyya reported presence of alkaloids, dihydrochelerythrine
and berberin, 15reported toddalinine, robustine, skimmianine, dictamnine, bergapten, luvangetin and isopimpinelline. Toddasin, Toddanol and Toddanone have been
isolated from T. asiatica16, 17, 18 have documented coumarins, fatty acid esters, β-sitosterol,
alkaloids and some other compounds. A new coumarin,
Toddalenone19 and an antiplasmodial coumarin were isolated from T. asiatica20.
Fruits are eaten for relief
from burning sensation in stomach21. Leaf and root are used to cure
rheumatic swellings and fever pain. The root bark is used medicinally as a
tonic and for the treatment of stomach ailments10, while the leaves
are used in patients having lung diseases or rheumatism and for the treatment
of fever, asthma and respiratory diseases10, 22. The alkaloids of
the crude extract have been shown to have anti-inflammatory effects in rats
using the carragennan test22 and to
inhibit the auricle swelling caused by xylol and
joint swelling caused by agar in rats24. T. asiatica are reported to have anti-malarial and
anti-leukimatic properties25, 26. The
central and peripheral antinociceptive effects of T.
asiatica have been demonstrated using mice27.
MATERIALS AND METHODS:
Plant collection
and authentication:
The plant specimens (Toddalia asiatica var. floribunda)
fruit were collected from Indira Nagar, Emerald, Udhagamandalam, The Niligiris, Tamil Nadu, India. Care
was taken to select healthy plants and normal organs.The plant material was
taxonomically identified and authenticated by Director, Plant Anatomy Research
Centre, Chennai A voucher specimen (PARC/250/07) has been deposited in the
herbarium Department of Botany, Periyar E.V.R.
College, Tiruchirappalli - 620 023, , Tamil Nadu,
India.
Pharmacognostical studies:
Organoleptic or Macroscopical character:
The fresh young
(green in colour) and mature fruit (Yellow in colour) were also subjected to
investigation. Studies, such as shape, size, outer surface, inner surface,
fracture, taste and odour of fruit, were carried out.
Microscopical studies:
Preparation of
specimen:
The required samples of
different organs were cut and removed from the plant and fixed in FAA (Farmalin-5ml+
Acetic acid-5ml + 70% Ethyl alcohol-90ml).After 24 hrs
of fixing, the specimens were dehydrated with graded series of tertiary -Butyl
alcohol as per the schedule13. Infiltration of the specimens was
carried by gradual addition of paraffin wax (melting point 58- 60o
C) until TBA solution attained super saturation. The specimens were cast into
paraffin blocks.
Sectioning:
The paraffin embedded
specimens were sectioned with the help of Rotary Microtome. The thickness of
the sections was 10-12 μm. Dewaxing of the sections was by customary procedure28.
The sections were stained with Toluidine blue as per the method15.
Since Toluidine blue is a polychromatic stain. The staining results were
remarkably good; and some cytochemical reactions were
also obtained. The dye rendered pink colour to the
cellulose walls, blue to the lignified cells, dark green to suberin,
violet to the mucilage, blue to the protein bodies etc. wherever necessary
sections were also stained with safranin and
Fast-green and IKI(for Starch). For studying the stomatal
morphology, venation pattern and trichome
distribution, paradermal sections (sections taken
parallel to the surface of leaf) as well as clearing of leaf with 5% sodium
hydroxide or epidermal peeling by partial maceration employing Jeffrey’s
maceration fluid28 were prepared.
Glycerine mounted temporary
preparations were made for macerated/cleared materials. Powdered materials of
different parts were cleared with Naoh and mounted in
glycerine medium after staining. Different cell component
were studied and measured29.
Photomicrographs:
Microscopic descriptions of
tissues are supplemented with micrographs wherever necessary. Photographs of
different magnifications were taken with Nikon labphoto
2 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.
Magnifications of the figures are indicated by the scale - bars.
Descriptive terms of the anatomical features are as given in the standard
Anatomy books30, 31.
FIG - 1: Fruit entire surface
view
PSg: Perisistent Stigma
FIG - 2: Cross Section view pf the fruit with seeds Pe: Pericarp; Sd: Seed
FIG - 3: T.S. of
fruit
LoC: Locule chamber; Pe:
Pericarp;
Se: Septal Partition; OV: Ovule
FIG- 4: Secretary Cavity
LoC: Locule chamber; Pe: Pericarp;
OV: Ovule; Pl: Placentum; Se: Septal Partition
RESULTS AND DISCUSSION:
Identification and
evaluation of plant drugs by pharmacognostical
studies is still more reliable, accurate and inexpensive. Standardization plays
an important role in the production of phytopharmaceuticals
of standard quality and purity
Microscopic investigation:
It is a liana with woody,
corky, thorny stems that climb on trees, rea- ching up to 10 meters in length. It has yellow-green
flowers, shiny green citrus-scented leaves, and orange fruits about half a
centimeter in diameter that tastes like orange peel.
In the present
study the fruit of Toddalia asiatica var. floribunda (fruit) was evaluated
for pharmacognostic analysis which revealed the following
results.
Anatomy of the fruit:
Anatomy of the fruit is sub globose, 3-6 celled, fleshy; Seeds are one in each cells (Fig: 1, 2). The fruit is a berry
with thick fleshy pericarp (Fig: 3, 4).
Locules:
The locules
are wide separated from each other by thick septa. The fruit consists of an
epidermis (exocarp), an inner thick, parenchymatous mesocarp (Fig: 3).
Exocarp:
The exocarp
includes two or three layers of tangentially stretched thick walled cells. There
are wide circular secretary cavities which originate by lysigenous
process of mesocarp cells. The cells surrounding
cavity are like epithelial cells. Some darkly stained spherical tannin bodies, are seen in and around the cavities (Fig: 5).
Fibres:
Small and large clusters of fibres’ and circular vascular bundles and tanninferous, Cells are seen in the pericarp (Figure: 6).
Macroscopic characteristics Macroscopic characteristics of Toddalia
asiatica var. floribunda (fruit) is shown
in Table 1.
Table1: Macroscopic
characteristics of Toddalia asiatica var. floribunda (fruit).
|
S. No. |
Parameters |
Observation of fruit |
|
1 |
Mature fruit - Yellow Color |
Young fruit - Green |
|
2 |
Odour |
Characteristic |
|
3 |
Taste |
Pungent |
|
4 |
Nature |
Coarse powder |
CONCLUSION:
After the present
investigation, it can be concluded that the Pharmacognostical
studies of the leaves and stem from Toddalia asiatica var. floribunda (fruit) yielded a set of
qualitative and quantitative arameters or standards
that can serve as an important source of information to ascertain the identity
and to determine the quality and purity of the plant materials for future
studies. These parameters also will serve as standard data for quality control
studies of pharmaceutical preparations from the leaves Toddalia asiatica var. floribunda
(fruit).
ACKNOWLEDGEMENT:
The author thanks Dr. P. Jayaraman, Director, Plant
Anatomy Research Centre, West Thambaram, Chennai for helping in the anatomical studies. Dr. M. Senthilkumar, Assistant Professor and Research Coordinator,
PG Research Department of Botany, Vivekanandha
College of Arts and Sciences for Women (Autonomous), Elayampalayam-
637 205, Tiruchengode, Tamil Nadu, India.
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Received on 04.07.2019
Modified on 31.07.2019
Accepted on 16.08.2019
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Res. J. Pharmacognosy and Phytochem. 2019; 11(3):193-196.
DOI: 10.5958/0975-4385.2019.00033.5