MICROSCOPICAL AND PHYSICO-CHEMICAL INVESTIGATIONS OF THE LEAVES OF PUTRANJIVA ROXBURGHII WALL.

MICROSCOPICAL AND PHYSICO-CHEMICAL INVESTIGATIONS OF THE LEAVES OF PUTRANJIVA ROXBURGHII WALL.

Madhavi R. Badole* and Vidya V. Dighe

Department of Chemistry, Ramnarain Ruia College, Matunga (East), Mumbai, Maharashtra, India

ABSTRACT

World Health Organization (WHO, Geneva) has appreciated the importance of medicinal plants for public health care for use in developing nations. Putranjiva roxburghii Wall. is having important role in the traditional Ayurvedic and Unani systems of holistic health and herbal medicine of the East. The leaves of Putranjiva roxburghii Wall. are reported to have good medicinal values in traditional system of medicine. The present study highlights the pharmacognostical studies including parameters such as powder analysis by microscopical evaluation, color reaction, and study of physicochemical parametersof the Putranjiva roxburghii Wall. leaves. These observations will help in the pharmacognostical identification and standardization of the drug in the crude form and can be used to distinguish the drug from its adulteration.

Physicochemical Parameters

INTRODUCTION:After decades of serious obsession with the modern medicinal system, people have started looking at the ancient healing systems like Ayurveda, Siddha and Unani. This is because of the adverse effects associated with the synthetic drugs. Herbal drugs play an important role in health care programs especially in developing countries ¹.

However a key obstacle, which has hindered the acceptance of the alternative medicines in the developed countries, is the lack of documentation and stringent quality control methods. There is a need for documentation of research work carried out on traditional medicines ².  

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 physicochemical studies.

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 medicines which will contribute to their safety and efficacy. Simple pharmacognostic techniques used in standardization of plant material include its morphological, anatomical and biochemical characteristics ³.

Putranjiva roxburghii Wall. is a moderate sized evergreen tree. The height is up to 12m with pendent branches and dark grey bark having horizontal lenticels. Leaves are simple, alternate, dark green, shiny, elliptic-oblong. Putranjiva roxburghii Wall. is found wild or cultivated in almost all parts of India ⁴.

Leaves of Putranjiva roxburghii Wall. are bitter, astringent, refrigerant and procreant. The leaves are useful in the treatment of catarrh, skin disease, fever and sterility. The leaves are given in decoction for cold and fever, and are also used in rheumatism ⁵.

The leaves of Putranjiva roxburghii Wall. posses analgesic, antipyretic and anti-inflammatory activity ⁶.No systematic studies have been reported for its pharmacognostical study.

Hence, an effort has been made to establish the pharmacognostical as well as physicochemical study of Putranjiva roxburghii Wall. leaves.

MATERIAL AND METHOD:

Plant material: Putranjiva roxburghii Wall. leaves were collected from Keshav shrushti, Mumbai, India. The leaves were washed with water to remove soil particles, dried in the shade, and finely powered. The powder was passed through the 85 mesh sieve and stored in an airtight container at room temperature (28 ± 2^oC).

Authentication: A herbarium of Putranjiva roxburghii Wall. was prepared and authenticated from Botanical Survey of India, Pune.

Powder analysis:

1. Microscopy: The powder of leaves of Putranjiva roxburghii Wall. was boiled with conc.HNO₃ to remove coloring matter and mounted on glass slide using  glycerin,  covered with cover slip and viewed under microscope. The powder was also stained with saffranin and examined under microscope.
2. Color Reactions: The powdered drug materials were treated with different chemical reagents so as to aid in detection of chemical constituents under ordinary day light by standard methods. To study the behavior of drugs a pinch of each drug was treated with different chemical reagents viz Conc. HCl, Conc. HNO₃, Conc. H₂SO_4, acetic acid, 5% NaOH solution, saturated solution of picric acid, 5% ferric chloride solution followed by 10N ammonia solution and colors were observed. The different colors obtained with these reagents are listed in Table 1.
3. Fluorescence behavior: To study the fluorescence nature of powder, a pinch of powder after bleaching with 5% chloral hydrate was treated with different chemical reagents viz. 1N NaOH in methanol, 1N NaOH in water, 50% HCl, 50% HNO₃, 50% H₂SO₄, acetic acid, petroleum ether, chloroform, picric acid, 5% ferric chloride solution, 5% iodine and methanol and observed under UV light. The different colors obtained with these reagents are listed in Table 2.

Physicochemical parameters: The determination of various physiochemical parameters such as total ash, acid insoluble ash, water soluble ash, moisture content and loss on drying were calculated as per Indian Pharmacopeia.

1. Total ash: About 2.0 g of dried powder of Putranjiva roxburghii Wall. were accurately weighed and transferred to different pre weighed silica crucibles and were ignited with a flame of Bunsen burner, for about 1 hour. The ignition was completed by keeping in muffle furnace, at 555⁰ C ± 20⁰ C, till a white carbon free ash was formed. The silica crucibles were then cooled in desiccators and weighed, and the result obtained is given in Table 3.
2. Acid insoluble ash: About 2.0 of dried powder of leaves of Putranjiva roxburghii Wall. was accurately weighed and transferred to different pre weighed silica crucible and was ignited with a Bunsen burner, for about 1 hour. The silica crucible was then kept in a muffle furnace at 555^oC ± 20^oC, till a white carbon free ash was formed. The ash obtained was moistened with concentrated HCl and evaporated to dryness, after which it was kept in an electric air oven, maintained at 135^oC ± 2^oC, for 3 hours. After cooling, the ash was taken in beaker and 25 mL of dilute hydrochloric acid (2N HCl) was added, and crucibles was kept covered and heated on a water bath, for 10 minutes.

It was allowed to cool, and contents was filtered through Whatman filter paper no. 41. The residue was then washed with water, till washings was free from chloride. The filter paper along with the residue of plant powder was placed in different silica crucible and ignited in a muffle furnace, at 550⁰ C ± 20⁰ C, for 1 hour. The crucible was cooled and weighed to a constant weight. The percent acid insoluble ash content was then calculated for dried powder of leaves of Putranjiva roxburghii Wall. and the result obtained is given in Table 3.

3. Water soluble ash: About 2.0 g of dried powder of leaves of Putranjiva roxburghii Wall.  was accurately weighed and transferred to a  pre weighed silica crucibles and was ignited with a Bunsen burner, for about 1 hour. The crucibles were then kept in a muffle furnance at 550^oC±20^oC, till a white carbon free ash was obtained. After cooling, the ash was taken in beaker and to it 2 mL of distilled water was added, and beaker was kept covered and heated on a water bath, for 10 min.

It was allowed to cool, and content was filtered through Whatman filter paper no. 41. The filter paper and the residue was placed in silica crucibles and ignited in a muffle furnace, at 55^oC ± 20^oC, for 1 hour. The crucible was cooled and weighed to a constant weight. The percent water soluble ash content was then calculated for dried powder of leaves of Putranjiva roxburghii Wall. and the result obtained is given in table 3.

4. Moisture content: About 100 mg of accurately weighed dried powder of leaf of Putranjiva roxburghii Wall. was transferred to the reaction vessel. Titration with Karl Fishcer reagent was carried out as described in Indian Pharmacopeia and the result obtained is given in table 3.

RESULTS AND DISCUSSION

Microscopic character: The dried leaf powder of Putranjiva roxburghii Wall. showed the presence of straight walled and polygon epidermal cells. (Fig. 1) The vessels have annular thickening (Fig. 2) The stomata is anomocytic i.e. the stoma is surrounded by varying number of cells.(Fig. 3) The plant showed the presence of unicellular as well as multicellular trichomes (Fig. 4, 5).

FIG. 1: EPIDERMAL CELL 40X

FIG. 2:  VESSEL 100X

FIG. 3: STOMATA 100 X

FIG. 4: UNICELLULAR TRICHOMES 40X

FIG. 5: MULTICELLULAR TRICHOME100X   

Behavior of Powder Drug with different Chemical Reagents: The powder drug with different chemical reagents shows different color when seen on naked eye and the color obtained with different reagents is given in Table 1, and the fluorescence observed with different reagent is given in Table 2.

TABLE 1: POWDER ANALYSIS WITH CHEMICAL AGENTS

TABLE 2: FLUORESCENCE ANALYSIS OF POWDERED DRUG

Physicochemical Parameters: The physicochemical parameters are important for identifying adulterants and improper handling of drugs. Table 3 reveals the result of physicochemical parameters of powdered drug, carried out by using standard procedures.

TABLE 3: PHYSICOCHEMICAL PARAMETERS OF LEAF POWDER OF PUTRANJIVA ROXBURGHII WALL.

CONCLUSION: The methods carried out in the present research work, like powder analysis and physico-chemical studies will serve as standard reference for identification and distinguishing Putranjiva roxburghii Wall. leaves powder from its substitutes and adulterants. This report would assist in the identification of the crude drug in future.

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   How to cite this article:

   Badole MR and Dighe VV. Microscopical and Physicochemical investigations of the leaves of Putranjiva roxburghii Wall. Int J Pharm Sci Res 2012; Vol. 3(8): 2599-2602.

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