Pharmacognostical Studies on Thespesia populnea Bark
R Parthasarathy1, R
Ilavarasan2, CM Karrunakaran1
1Bharath
University, Selaiyur, Chennai-600 073, Tamil Nadu.
2Captain Srinivasa Murthi Drug Research Institute for Ayurveda
and Siddha, Chennai-600 109, Tamil Nadu.
Thespesia Populnea is a reputed ever green tree
belonging to the family malvaceae; 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 astringent, antibacterial, hepatoprotective, haemostatic, anti-diarroheal
and anti-inflammatory. 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,
physical evaluation and preliminary phytochemical
screening of various successive extracts were carried out and the parameters
were reported. These studies indicated the possible information for correct
identification and standardization of this plant material.
KEYWORDS: Thespesia populnea,
bark macroscopy, bark microscopy.
INTRODUCTION:
Thespesia populnea soland ex correa
(family malvaceae) is a large tree found in the
tropical regions and coastal forests in India and cultivated in the gardens.
All the parts of the plant used in traditional system of medicine. The bark,
leaves, flower and fruits are useful in cutaneous
infection such as scabies, psoriasis, eczema, ringworm, and guinea worm. The
decoction of the bark is commonly used for the treatment of skin and liver
diseases. A compound oil of bark and capsules is useful in urethritits
and gonorrhea. The bark, root, fruits were used in dysentery, cholera and
hemorrhoids1. The fruits of
the plant are used in ayurveda for the control of
diabetes2. The barks and flowers posses
astringent, hepatoprotective, antioxidant and
anti-inflammatory activities in rats3,-6.
The leaves and bark
of this tree are still used to produce oil for the treatment of fracture wounds
and as an anti-inflammatory poultice applied to ulcers and boils, as a folk
medicine6. Gossypol was found to be the major component of Thespesia populnea7 producing
anti-fertility effects in rats8,9 as well
as in human beings10. Four naturally occurring quinones
viz thespone, thespesone, mansonone-D, and mansonone-H have been extracted from heart wood of the
plant11. The phytochemical study of bark
reveals the presence of gossypol, tannin and coloring matter12.
Lack
of proper standard of medicinal plants may lead to usage of substandard drugs
which will cause damage to the faith on traditional system of medicine.
Therefore scientific method must be developed to identify and maintain quality
of plant drugs. With this aim the present investigation was planned to study
the pharmacognostical aspects of Thespesia
populnea bark.
.
Table 1: Loss on drying and ash
values of powdered stem bark of Thespesia populnea
Parameters
|
Result |
|
Loss
on drying |
5.46 |
|
Ash
values 1)
Total ash |
4.72 |
|
2)
Acid insoluble ash |
0.96 |
|
3)
Water soluble ash |
3.42 |
FIG-1: TS of Thespesia populnea bark.
Co: cortex; Ph: pholem; Fi: fissures; Pe: periderm; DR: dilated rays.
MATERIALS AND METHODS:
Plant materials:
The
fresh bark from about 3-5 years old tree were collected from our Salaiyur University campus in Chennai, India. The plant
material was taxonomically identified and authenticated by Director, Plant
Anatomy Research Centre, Chennai
A
voucher specimen (PARC/236/07) has been deposited in the herbarium of the same
department. Then the stem barks were shade dried at temperature 25-300.
Pharmacognostical studies:
Organoleptic or Macroscopical character:
The
dried stem barks were also subjected to investigation. Studies, such as shape,
size, outer surface, inner surface, fracture, taste and odour
of bark, were carried out. A yellow exudates was oozed
out from the fracture surface.
Anatomical studies or Microscopical
character:
Preparation of specimen:
The
bark were cut into required size and fixed in FAA (Formalin 5ml + Acetic acid 5
ml + 70% Ethanol 90 ml). After 24 hrs of fixing, the specimens were dehydrated
with graded series of tertiary butyl alcohol. Infiltration of the specimens was
carried by gradual addition of paraffin wax (melting point 58-600C)
until tertiary butyl alcohol solution attained super saturation. The specimens
were cased into paraffin blocks.
Sectioning:
The
paraffin embedded specimen was sectioned with the help of rotary microtome. The
thickness of the section was 10-12 µm. After dewaxing
the sections were stained with toluidine blue. Since toluidine blue is a polychromatic stain, the staining
results were remarkably good and some phytochemical
reactions were obtained. The dye rendered pink color 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 also stained with safranin and fast green
and iodine (For starch)13,14.
FIG-2: TLS of Thespesia populnea bark.
Photomicrographs:
Microscopic
descriptions of tissues are supplemented with micrographs wherever necessary.
Photographs of different magnifications were taken with Nikon Labphot-2
microscope units. For normal observations bright fields was used. For the study
of crystals, starch grains and lignified cells polarized light were employed.
Since these structures have birefringent property,
under polarized light they appear bright against dark back ground.
Physico-chemical evaluation:
Physical
parameters such as loss on drying, total ash, acid insoluble ash, water soluble
ash were determined as per the Indian Pharmacopeia15. All the
results were recorded carefully (Table no: 1).
Phytochemical screening:
The dried and powdered stem
bark was subjected to preliminary phytochemical
screening for qualitative detection of phytoconstituents.
The dried and coarsely powdered stem bark (100 g) was extracted successively
with petroleum ether (40-60ºC), chloroform (59.5-60ºC), ethyl acetate
(76.5-77.5ºC), and ethanol (90%) in a soxhlet
extractor by continuous hot percolation. Finally the marc was macerated with
chloroform water. Each time before extracting with the next solvent of higher
polarity the powdered drug (marc) was dried in a hot air oven below 50ºC for 10
minutes. Each extract was concentrated by distilling off the solvent, which was
recovered subsequently. The concentrated extracts were evaporated to dryness
and the extracts obtained with each solvent were weighed. Their percentages
were calculated in terms of initial air dried plant material. They are
tabulated (Table no: 2)16.
Table 2: Results of phytochemical screenings of successive extracts of stem
bark of Thespesia
populnea
|
Constituent |
Pet. Ether extract |
Chloroform extract |
Ethyl acetate
extract |
Ethanol extract |
Aqueous extract |
|
Alkaloids |
- |
- |
- |
- |
- |
|
Carbohydrates |
- |
- |
- |
+ |
+ |
|
Glycosides |
- |
- |
- |
+ |
+ |
|
Steroids |
- |
- |
- |
- |
- |
|
Flavonoids |
- |
- |
- |
+ |
- |
|
Saponins |
- |
- |
- |
- |
- |
|
Fixed oils and fats |
- |
- |
- |
- |
- |
|
Tannins |
- |
- |
- |
+ |
+ |
|
Proteins and amino
acids |
- |
- |
- |
- |
+ |
|
Mucilage |
- |
- |
- |
- |
+ |
Table 3: Percent extractives of
successive extracts of Thespesia populnea stem bark
|
Solvent |
Extractive
values (% w/w) |
|
Pet.
Ether |
2.52 |
|
Chloroform |
4.18 |
|
Ethyl
acetate |
2.72 |
|
Ethanol |
12.50 |
|
Water |
10.84 |
RESULT AND DISCUSSION:
Macroscopical characters:
The
bark of Thespesia populnea
were varies from 5-10 cm long, 3-5 cm width and 0.6-0.10 cm thick. The outer
surfaces were dull brown or grey in color, rough due to longitudinal and
transverse crack and fissure were seen. Exfoliation of the outer bark was seen
in some place. Inner surface were striated and pale yellowish in color. The
fractures were short and fibrous. The odour and taste
were aromatic, characteristic and slightly bitter. A yellow exudates was oozed
out from the fractured surface probably mucilage
Microscopical characters:
Transverse
section of the bark showed the following distinct characters:
Periderm: The stem of the epidermis were replaced by a broad
zone of periderm. The periderm
comprises of five to eight layers of suberized cork
cells and three or four layers of thin walled phelloderm
cells. Fissures of various shapes and wide lenticels were seen in the periderm.
Cortex: The periderm is followed by a
narrow cortical zone which consists of several layers of tangential oblong
compact parenchyma cells.
Phloem: It consists of wide secondary phloem comprises of radial series of phloem
elements alternating with tangential blocks of phloem fibers. Funnel shaped
dilating rays in between the radial cones of phloem. This zone is
differentiated into outer collapsed phloem and inner intact phloem. The
collapsed phloem has wide, funnel shaped, dilating rays alternating with radially extended, conical phloem fibers and crushed sieve
elements. The dilating rays have tangential rows of narrowly rectangular parenchyma
cells. The radial, conical segments of phloem have thick bands of phloem fibers
alternating with crushed phloem elements (Fig: 1).
TLS
view of the phloem: In tangential
longitudinal section the structure of the phloem rays and sieve elements were
seen. The phloem rays are non storied. They are wide
and very high. The rays range from uniseriate to multiseriate. The multiseriate
rays are three or four cells thick. There are homocellular
consisting of only one type of cells namely polygonal, thin walled cells. The
rays range in height from 140-750 µm with average
of 490 µm.
The uniseriate and biseriate
rays are 90 to 170 µm within on average 130 µm. Mucilage cavities are seen
frequently in the phloem zone. The sieve tube members are long and narrow. They
have simple, oblique sieve plate. The phloem parenchyma cells are fusiform type and they appear to be storied. Calcium
oxalate druses are abundant in the medullary ray
cells. They occur mostly in singles (Fig: 2,3).
For identification and
evaluation of plant drugs by pharmacognostical
studies is still more reliable, accurate and inexpensive17. The macroscopical studies of bark revealed the presence of
characteristic aromatic odour, color, taste and the
fracture surface. The microscopical study indicated
the presence of a typical arrangement of phloem tissues, mucilage cavity, medullary rays and the absence of sclerides, etc may be useful
for their identification.
In physiochemical
studies of various parameters established, like various ashes content which
showed the presence of inorganic salts by naturally occurring or adhering to
it, or deliberately added to it as a form of adulteration. These values are
important quantitative standards. The various extractive values obtained by
results showed higher yield in ethanolic extract14.
The extracts
obtained by exhausting plant materials with specific solvents are indicative of
approximate measures of their chemical constituents extracted with those
solvents from a specific amount of air-dried plant material. This parameter is
employed for materials for which as yet no suitable chemical or biological
assay exists. The results also showed higher extractive values in
hot extraction, indicating the effect of elevated temperature on extraction.
In all methods alcohol yielded higher extractives18.
The plant
material was subjected to preliminary phytochemical
screening involving successive solvent extraction by different solvents in
order of increasing polarity to obtain diverse polar and non polar phytoconstituents possessing different solubility pattern,
followed by various chemical tests for qualitative detection of various
chemical constituents. The percent extractives in different solvents indicate
the quantity and nature of constituents in the extract19.
CONCLUSION:
The present
investigation it can be concluded that the pharmacognostical
study of Thespesia populnea
bark yielded a qualitative and quantitative parameters or standards that can
serve as an important possible sources of information to ascertain the identity
and to determine the quality and purity of the plant material.
These
information will also be helpful to differentiate Thespesia
populnea from the closely related other species
and varieties of Thespesia.
FIG-3: Crystal distribution in TLS of Thespesia populnea bark.
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Received on 07.06.2009
Accepted on 14.08.2009
© A&V Publication all right reserved
Research Journal of Pharmacognosy and Phytochemistry.
1(2): Sept. - Oct. 2009, 128-131