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Indigenous systems of medicine practices in India are based mainly on the use of plants. Standardization of plant material to be used as medicine can be achieved by stepwise pharmacognostic and phytochemical studies.
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Phytochemical Pharmacognostical and Physicochemical Standardization of Peperomia Pellucida (l.) Hbk.
Indigenous systems of medicine practices in India are based mainly on the use of plants. Standardization of plant material to be used as medicine can be achieved by stepwise pharmacognostic and phytochemical studies.
Indigenous systems of medicine practices in India are based mainly on the use of plants. Standardization of plant material to be used as medicine can be achieved by stepwise pharmacognostic and phytochemical studies.
Majumder P / Pharmacie Globale (IJCP) 2011, 8 (06)
1 Pharmacie Globale
(IJCP), Vol. 02, Issue 08
Available online at www.pharmacie-globale.info
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
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 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. Research Article
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
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) 3 Pharmacie Globale
(IJCP), Vol. 02, Issue 08
Figure 4. Powder characteristics study of P. pellucida stem (X450).
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 calcium oxalate are scattered throughout the ground tissue.; F, unicellular trichomes. Physicochemical evaluation The Loss on Drying (LOD), Ash Values likes (Total Ash, Acid insoluble ash, Water soluble ash) Water soluble extractive, alcohol soluble extractive, foaming index, Swelling Index and measurement of fibers of stem powder are given in table 2. Table 2. Quality control parameters of Peperomia pellucida stem. Sl no Parameter Value; % 1 Ash values Total ash 0.490.02 Water soluble ash 0.2740.01 Acid insoluble ash 0.0250.01 2 Extractive values Ethanol (90%) 7.250.02 Aqueous 15.050.03 3 Loss on drying Dry matter content 82.70.01 Moisture content 17.30.15 4 Foaming index Nil 5 Swelling index 0.80.03 6 Fibers (in ) Avg. Length 24.030.23 Avg. Width 150.610.41 N= Three experiments for each parameter. Values shown are mean S.E. Phytochemical Screening 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 - 5 Tannins + 6 Flavonoids + 7 Steroids + 8 Triterpenoids +
Powder Drug with different Chemical Reagents The powder drug with different chemical reagents show different colour when seen with naked eye as shown in Table 4. Fluorescence Analysis Peperomia pellucida stem and stem powder produced different colour with different chemical reagents under UV light (short wavelength; 254nm and long wavelength; 356nm). The fluorescence characteristics of powdered drug are shown in table 5. Table 4. Colour characters of plant powder in different solvents Sl no. Drug treatment Colour 1 Powder as such Pale green 2 Powder + Picric acid Yellowish green 3 Powder + 50% HNO3 Orange 4 Powder + 1N HCl Dark Brown 5 Powder + 50%H2SO4 Brown 6 Powder + Glacial acetic acid Yellowish green 7 Powder +10% FeCl3 Green 8 Powder + Iodine Brown Table 5. Fluorescence characters of stem powder in different solvents Sl no Particulars of treatment Under ordinary white light Under UV 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 Establishing standards is an integral part of establishing the correct identity and quality of a crude drug. Before any drug can be included in the pharmacopoeia, these standards must be established. The majority of the information on the identity, purity and quality of the plant material can be obtained from its macroscopy, microscopy and physicochemical parameters. As there is no record on pharmacognostical work on stems of Peperomia pellucida, The present work is undertaken to produce some pharmacognostical standards and this foundings may help to proper identification and ensures the quality of the drug and also help this amazing plant grown on commercial basis for better use in pharmaceutical herbal formulations.
ACKNOWLEDGEMENT The author is thankful to the honorable chairman of Rajiv Gandhi Educational Trust and also grateful to the Principal, Rajiv Gandhi Institute of Pharmacy, Trikaripur, Kasaragod (Dist), Kerala, for giving all encouragement and valuable support to carry out this research work.
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