Majumder P / Pharmacie Globale (IJCP) 2011, 8 (06)

Available online at www.pharmacie-globale.info ISSN 0976-8157

Research Article
PHARMACIE GLOBALE
INTERNATIONAL JOURNAL OF COMPREHENSIVE PHARMACY

PHYTOCHEMICAL, PHARMACOGNOSTICAL AND PHYSICOCHEMICAL STANDARDIZATION

OF Peperomia pellucida (L.) HBK. STEM
Pulak Majumder
Department of Pharmacognosy, Rajiv Gandhi Institute of Pharmacy, Trikaripur, Kasaragod, Kerala, India.
Received: 12 June 2011; Revised: 22 July 2011; Accepted: 29 July 2011; Available online: 7 August 2011

ABSTRACT
Peperomia pellucida is an America and Asia originated small herb belonging to the Family Piperaceae. The stems
of Peperomia pellucida are reported to have good medicinal values in traditional system of medicine. The stems of
Peperomia pellucida were collected locally, shade dried and extracted with methanol and water by using Soxhlet
apparatus. The yield of methanolic and water extracts of stem were 7.25% and 15.05% respectively. The
preliminary phytochemical screening was carried out for the presence of Carbohydrates, Alkaloids, Tannins,
Flavonoids, Steroids, Triterpenoids etc and absence of saponins and proteins for methanolic extracts of
Peperomia pellucida (stems). The physical evaluation was carried out for the determination of methanol-soluble
extractive value, water-soluble extractive values; ash value includes total ash, acid insoluble ash and water-
soluble ash, foaming index, swelling index, fibres measurement and moisture content for stems of Peperomia
pellucida. The present study also highlights the Pharmacognostical studies on the stems of the plant Peperomia
pellucida. These observations will help in the Pharmacognostical identification and standardization of the drug in
the crude form and also to distinguish the drug from its adulteration.
Keywords: Peperomia pellucida, Microscopy, Phytochemical screening, Physical evaluation.

INTRODUCTION
The knowledge about the use of medicinal plants has been
accrued through centuries and such plants are still valued
even today, although synthetics, antibiotics etc have
attained greater prominence in modern medicine. The
indigenous systems of medicine practices in India are
based mainly on the use of plants.1 Herbal drugs play an
important role in health care programs especially in
developing countries. Ancient Indian literature
incorporates a remarkably broad definition of medicinal
plants and considers ‘all’ plant parts to be potential
sources of medicinal substances.2 Unfortunately the use of
medicinal plants in these countries is based primarily on
empirical knowledge, and many of the plants have not
been scientifically evaluated for their safety and efficacy.3
There is a need for documentation of research work
carried out on traditional medicines.4 With this backdrop,
it becomes extremely important to make an effort towards
standardization of the plant material to be used as
medicine. The process of standardization can be achieved
by stepwise pharmacognostic and phytochemical studies.5
These studies help in identification and authentication of
the plant material. Correct Identification and quality
assurance of the starting materials is an essential
prerequisite to ensure reproducible quality of herbal
medicine which will contribute to its safety and efficacy.
Simple pharmacognostic techniques used in
standardization of plant material include its
*Corresponding Author:
Pulak Majumder
Lecturer, Department of Pharmacognosy,
Rajiv Gandhi Institute of Pharmacy, Trikaripur, Kasaragod, Kerala, India.
Contact no: +91-7736404410; Email: pulak2007@gmail.com
morphological, anatomical and biochemical
characteristics.6
The genus Peperomia pellucida is a member of the family
Piperaceae. Peperomia pellucida is a common fleshy
tropical annual, shallow-rooted herb, usually growing to a
height of about 15 to 45 cm. It is characterized by fibrous
roots, succulent stems, shiny, heart-shaped, fleshy leaves
and tiny, dot-like seeds attached to several fruiting spikes.
It has a mustard-like odor when crushed (Figure 1).
Figure 1. Plant of Peperomia pellucida
The plant has a thread like but angular trailing stem.
Those growing in rich habitats have fleshy and stout
stems. They are translucent pale green, erect or ascending,
usually 15-45 cm long, internodes usually 3-8 cm long and
glabrous. Leaves are alternate, blunt, heart shaped and as
transparent and smooth as candle wax grows as a long
shrubby looking creeping cover or as an epiphyte. They
are medium green on upper surface, lower surface whitish
green, thinly fleshy, drying papery, broadly ovate, 1.5- 4 (-
5) cm long, 1-3.3 cm wide, palmately 3-nerved or 5-
nerved, glabrous, apex acuminate, base subcordate to
truncate, petioles 0.5-2 (-3) cm long, glabrous. The

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 Majumder P / Pharmacie Globale (IJCP) 2011, 8 (06)
elongated stems look like a vine with leaves rising 6 to 9
cm above the surface. Both leaves and stems have shiny
waxy surfaces. The foliage of the plant looks ornamental.
Flowers are very small, well-spaced, unnoticeable and bi-
sexual growing in the form of cord-like spikes arising from
the leaf axils, 1 to several, terminal and axillary or leaf-
opposed.
Medicinally in the South America, solution of the fresh
juice of stem and leaves is used against eye inflammation.7
Infusion and decoction of leaves and stems are used for
gout and arthritis. It is described in Ayurveda as: Rasa –
Katu and Madhur; Guna- Lakhu, rooksha, Teekshna; and
Virya- Ushna. The plant is described to passify vitiated
cough, pitta, constipation, kidney diseases, urinary
retention, dysuria, urinary tract infections, emaciation,
edema and general weakness. Infusion and decoction of
leaves and stems of fresh plant are eaten as salad for the
treatment of gout and arthritis.8,9
No systematic studies have been reported for its
pharmacognostical and phytochemical study hence an
effort has been made to establish the Pharmacognostical
as well as phytochemical study of Peperomia pellucida
stems.
MATERIALS AND METHODS
Plant Material
Stems of P. pellucida were collected from the Trikaripur
forest areas, kasaragod district of Kerala, India, in the
month of November 2010 in a quantity sufficient for all
the experiments in a single batch. The plant material was
authenticated by Dr. Khaleel. Course director, Dept. of
Environmental Studies, Kannur University, Payyanur and
specimen was submitted and preserved in the Department
of Pharmacognosy Rajiv Gandhi Institute of Pharmacy (No.
RGIP/Cog/10-11/01), Trikaripur.
Preparation of the stem extract
The stem of plant were dried under shade, separated and
made to dry powder. It was then passed through the 40
mesh sieve. Dried and powered plant defatted firstly to
remove fatty material for this purpose 150 g of weighed
powered plant of Peperomia pellucida was packed in
Soxhlet apparatus and extracted with methanol (90%) and
there after distilled water for 36 hrs and completion of
extraction was confirmed by pouring a drop of extract
from the thimble on a filter paper, which does not show
the presence of any oil spot on that. After complete
extraction the solvent was evaporated and concentrated to
dry residue. The methanol and aqueous extract of
Peperomia pellucida stems yielded greenish brown and
deep brown semi solid residue respectively.
Pharmacognostic studies
Morphology of fresh stems of P. pellucida was studied.
Photomicrography of unstained transverse sections and
stained transverse sections (using phloroglucinol-HCl) of
fresh stem was performed.10 The stems were dried under
shade, powdered, stored in airtight containers and used
for powder study, physico-chemical evaluation and
phytochemical screening.
Physicochemical evaluation
The crude plant material was subjected to the physical
evaluation. The various parameters was evaluated such as
solvent extractive value, its includes water soluble,
methanol soluble extractive value, moisture content, ash
value including acid insoluble and water soluble ash,
foaming index and swelling index.11,12
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Phytochemical screening
The Phytochemical examination of the methanolic extract
was performed by the standard methods and shows the
presence of various phytochemical constituents.13-17
Powder Drug with different Chemical Reagents
The powder drug with different chemical reagents show
different colour when seen on naked eye.
Fluorescence Analysis
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 UV light. The fluorescence characteristics of
powdered drug were studied under UV light (254nm and
356nm) after treating with different chemical reagents.
RESULTS AND DISCUSSION
Macroscopy and Microscopy
Stems are succulent, translucent pale green, erect or
ascending, internodes usually 3-8 cm long, glabrous and
hairless (Table 1, Figure 2).
Table 1. Macroscopic evaluation of P. Pellucida stem.
Character When fresh After drying Powder
Color Yellowish green Green to yellowish Gray
Odor Caracteristic Caracteristic Caracteristic
Taste Acrid Acrid Acrid
Texture Thin soft Fibrus Fibrus
Fracture Pale green Straw green Dark green
Figure 2. P. pellucida dried stem
Microscopically, the fresh plant transverse section of the
stem has shown a polystelic structure. Each collateral
vascular bundle and fascicular cambium is covered with a
uniseriate parenchymatous pericycle and an endodermis
with casperin strips. The epidermis is unilayered with
cutical, secretory trichroms and periclinal thick walled
cells. The cortex possesses parenchyma and subepidermic
collenchyma arranged in irregular strate. Endodermis
contains 2-4 vascular bundles (Figure 3).
Figure 3. Microscopic evaluation of P. Pellucida stem.
Stem structure in cross-section shows overall diagram detail and vascular
bundle of P. pellucida (CL = collenchyma, EP = epidermis, FA = fibers; PA =
parenchyma, VB = vascular bundle, XY=xylem, PH= phloem).
Powder characteristics P. pellucida stem.
The powder microscopy of the stem powder of Peperomia
pellucida showed the presence of cork cells 2 to 3 layers,
round shaped oil glands, sleder fibres, starch grains,
vessels accociated with fibres, tricromes, xylem cells,
calcium oxalate crystals etc. (Figure 4).
 Majumder P / Pharmacie Globale (IJCP) 2011, 8 (06)

Figure 4. Powder characteristics study of P. pellucida Phytochemical Screening

stem (X450). The result of preliminary phytochemical screening of the
methanolic extract of Peperomia pellucida revealed the
presence of alkaloids, carbohydrates, terpenoids, tannins,
Flavonoids, steroids and absence of saponins and protein.
(Table 3)
Table 3. Preliminary phytochemical analysis of
stem extract of P. Pellucida.
Sl no Phyto- Constituents Stem extract
1 Carbohydrates +
2 Proteins -
3 Alkaloids +
4 Saponins -
A, Xylem vessels with reticulate thickening; B, Orange matter; C, Cork cells; D,
Pericyclic fibers; E, Calcium oxalate cluster crystal Starch grains and prisms of 5 Tannins +
calcium oxalate are scattered throughout the ground tissue.; F, unicellular 6 Flavonoids +
trichomes.
7 Steroids +
Physicochemical evaluation 8 Triterpenoids +
The Loss on Drying (LOD), Ash Values likes (Total Ash,
Acid insoluble ash, Water soluble ash) Water soluble Powder Drug with different Chemical Reagents
extractive, alcohol soluble extractive, foaming index, The powder drug with different chemical reagents show
Swelling Index and measurement of fibers of stem powder different colour when seen with naked eye as shown in
are given in table 2. Table 4.
Table 2. Quality control parameters of Peperomia Fluorescence Analysis
pellucida stem. Peperomia pellucida stem and stem powder produced
Sl no Parameter Value; % different colour with different chemical reagents under UV
1 Ash values light (short wavelength; 254nm and long wavelength;
Total ash 0.49±0.02 356nm). The fluorescence characteristics of powdered
Water soluble ash 0.274±0.01
drug are shown in table 5.
Acid insoluble ash 0.025±0.01
2 Extractive values
Table 4. Colour characters of plant powder in
Ethanol (90%) 7.25±0.02
different solvents
Sl no. Drug treatment Colour
Aqueous 15.05±0.03
3 Loss on drying 1 Powder as such Pale green
Dry matter content 82.7±0.01 2 Powder + Picric acid Yellowish green
Moisture content 17.3±0.15 3 Powder + 50% HNO3 Orange
4 Foaming index Nil 4 Powder + 1N HCl Dark Brown
5 Swelling index 0.8±0.03 5 Powder + 50%H2SO4 Brown
6 Fibers (in µ) 6 Powder + Glacial acetic acid Yellowish green
Avg. Length 24.03±0.23 7 Powder +10% FeCl3 Green
Avg. Width 150.61±0.41 8 Powder + Iodine Brown
N= Three experiments for each parameter. Values shown are mean ± S.E.
Table 5. Fluorescence characters of stem powder in different solvents
Under UV light
Sl no Particulars of treatment Under ordinary white light
Short Wavelength (254nm) Long Wavelength (366nm)
1 Stem as such Greenish yellow Black Grayish brown
2 Stem Powder as such Gray Grayish yellow Greenish white
3 Powder + 50% H2SO4 Black Green Green
4 Powder + 1N HCl Brown Greenish black Green
5 Powder + 50% HNO3 Golden yellow Black Light green
6 Powder + 5% KOH Greenish black Greenish black Green
7 Powder +MeOH Dark brown Brown Light green

CONCLUSION may help to proper identification and ensures the quality

Establishing standards is an integral part of establishing of the drug and also help this amazing plant grown on
the correct identity and quality of a crude drug. Before any commercial basis for better use in pharmaceutical herbal
drug can be included in the pharmacopoeia, these formulations.
standards must be established. The majority of the
ACKNOWLEDGEMENT
information on the identity, purity and quality of the plant
The author is thankful to the honorable chairman of Rajiv
material can be obtained from its macroscopy, microscopy
Gandhi Educational Trust and also grateful to the
and physico–chemical parameters. As there is no record
Principal, Rajiv Gandhi Institute of Pharmacy, Trikaripur,
on pharmacognostical work on stems of Peperomia
Kasaragod (Dist), Kerala, for giving all encouragement and
pellucida, The present work is undertaken to produce
valuable support to carry out this research work.
some pharmacognostical standards and this founding’s
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