INTRODUCTION:

Ficus nervosa Heyne ex Roth is a medium sized evergreen tree belongs to Moraceae family. Vernacular names of the tree are Vonjari in Telugu, Neer-al in Tamil, Eechamaram in Malayam and Narrowed leaved fig in English. It occurs in India, Australia, China, Taiwan, Malaysia, rare along streams in valleys of deciduous forests, Srivarimettu, Talakona, Tirumala, East- Godavari, Visakapatnam in Andhra Pradesh (Madavachetty et al., 2008; Gamble, 1967). Aerial roots are absent; leaves arecoriacuous, glabrous on both sides, oblanceolate to oblong 8-20 cm long, entire margin, narrowed at base. Ficus nervosa, is traditionally using for pain and inflammation in and around thirumal hills. (Madhava Chetty et al., 2008). Bark is brown mottled white, wood is white in colour and soft (Gamble, 1967). Therefore, the present investigation was planned to study the pharmacognostical and phytochemical aspects of Ficus nervosa Heyne ex Roth.

MATERIALS AND METHODS:

Collection and Authentication

The leaves of Ficus nervosa Henye ex Roth (Moraceae) were collected from Tirumala Hills, Tirupathi, India and it was identified and authenticated. The taxonomical identification and authentication of the plant was done by Dr. K. Madhava Chetty, Professor, Department of Botany, Sri Venkateswara University, Tirupati. The voucher specimen was preserved in our laboratory for further reference.

Morphological and Microscopical Investigation

Macroscopical examination was carried out to the freshly collected leaves and to the powder. In these tests colour, odour, taste, size and shape of leaf and powder were observed and noted and photographs were taken in the original environment.

Microscopic Studies

The fresh sample were cut into small pieces and fixed in FAA solution (Formalin 5ml + Glacial acetic acid 5ml + 70% Ethanol 90ml). After fixing, the specimens were dehydrated with graded series of tertiary butyl alcohol (TBA) as per the standard procedure (Sass, 1940). After complete dehydration, the specimens were embedded in paraffin wax. The paraffin embedded specimens were sectioned with the help of Rotary microtome (thickness 10-12µm). Dewaxing and staining of the sections were done by customary procedure. Sections were stained mostly with toluidine blue (Johansen, 1940). For anatomical studies the following staining schedules were followed.Tannic Acid – Ferric Chloride counterstained with 0.5% alcoholic safrain. This schedule was found to be quite satisfactory for all young plant tissues in which the primary walls were stained. Alcoholic safrain (0.5%) counterstained with 0.25% fast green. This schedule gives good result for studying the histology of different tissues of the plant organs especially the cell inclusions. Toludine Blue – O stain was prepared by dissolving 0.25g of the stain in the mixture of benzoic acid 0.25g, sodium benzoate 0.29g and distilled water 200ml with p^Hof 4.2 – 4.4. Since Toluidine blue is a polychromatic stain, the staining results were remarkably good and the dye render 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 (O’Brien, 1964). After dewaxing, the slides were stained for 5 – 10 minutes and then dehydrated. All permanent slides, after staining were dehydrated by using graded series of Ethanol + Xylol and mounted in DPX. Photomicrographs were done on Nikon–Labphot – 2 microscope using Konica colour film (100 ASA). For normal observations bright field was used. For the study of crystals and starch grains, the sections were photographed under polarized light. Magnifications of the figures are indicated by scale bars. Descriptive terms of various observations are as found in standard Anatomy books (Esau, 1979).

Physico chemical Parameters:

Physical constants of crude drugs like moisture content, ash values, extractive values and fluorescence analysis were determined by using official methods (Indian pharmacopeia-1996, Chase and Pratt 1949, Kokoshi et al., 1958 and Ushakumari et al., 2004).

RESULTS AND DISCUSSION:

Morphological and Microscopical Investigation

Morphological as well as microscopical studies of plants are the primary steps to establish its botanical standards before going to other studies. Morphological examination of leaf reveals the following interpretations: The plant Ficus nervosa is a moderate to large tree grows up to 35m tall. The colour of the leaves are Pale green, it has characteristic odour, with bitter to astringent taste. Size of the leaf is 8-20 cm in length, coriaceous shape, acute apex, narrowed base and entire margin.

Microscopical studies of the leaf showed the following:

The leaf is dorsiventral with distinct differentiation of the mesophyll tissue and smooth surfaces. The lamina is 240µm thick. The adaxial epidermis is fairly thick and the cells are rectangular to square shaped. The abaxial epidermis is slightly thinner and the cells are tubular in shape. Palisade mesophyll is present and the spongy parenchyma observed at the lower part of lamina. The life margin is blunt and semicircular and slightly bent. The midrib is quite prominent, projecting much beyond the surface of lamina. The vascular system of midrib consists of shallow wide “abaxial arc” and narrow “adaxial arc” of xylem and phloem. Calcium carbonate crystals of cystoliths are spherical bodies with spiny surface. They have fairly long, slender stalk with which they are attached to the roof of a wide chamber known as “litho cysts”. Epidermal cells appear polygonal in outline. Stomata are densely distributed on the abaxial side of the lamina. The stomata are “Actinocytic” type, that is, the stomata is surrounded by a radiating circle of 5-7 subsidiary cells. The lateral veins and veinlets gradually decrease in size and form dense reticular venation. The vein-islets are fairly distinct, they are rectangular to polygonal in outline. The vein-terminations are less prominent. When distinct they are long and slender, simple or branched.

T.S. of lamina of leaf

T.S. of leaf –margin

T.S. of leaf through midrib with lamina

T.S. of midrib

T.S. of lamina showing cystolith

Stomata of leaf

Cleared leaf showing vein-islet and vein termination

Physico-Chemical Parameters:

Table 1: Physico-Chemical parameters of powdered leaves of Ficus nervosa Heyne ex Roth

S. No.	Parameters	Average % W/W
1.	Ash values a) Total ash b) Acid insoluble ash c) Water soluble ash	7.13 1.14 1.41
2.	Extractive values a) Alcohol soluble Extractive b) Water soluble extractive	1.08 1.10
3.	Moisture content Loss on drying	2.14

Fluorescence Analysis:

Table 2: Fluorescence analysis of powdered leaves of Ficus nervosa Heyne ex Roth (Moraceae)

Treatments	Observations
Treatments	Day light	Long UV	Short UV
Powder as such	green	Light green	Green
Powder + 1N NaOH (aqueous)	Orange	Green Fluorescence	Brown
Powder + 1N NaOH (alcoholic)	Green	Yellow	Brown
Powder + 1N H₂SO₄	Yellow	Yellow Fluorescence	Light green
Powder + 1N HNO₃	Pale yellow	Light green	Pale green
Powder + 1N HCl	Green	Brownish	Orange
Powder + Ammonia	Brown	Dark green	Green
Powder + Acetic acid	Light green	Brown	Green Fluorescence
Powder + Iodine	Pale yellow	Green	Light green
Powder + FeCl₃	Light green	Dark green	Yellowish
Powder + water	Pale yellow	Pale yellow	Light green

Table 3: Fluorescent analysis of various extracts of Ficus nervosa Heyne ex Roth (Moraceae)

Extract	Day light	Long UV	Short UV
Petroleum ether	Light green	Yellow	Green
Chloroform	Green	Bluish green	Green fluorescence
Methanol	Green	Green Fluorescence	Green
Distilled water	Brown	Light Green	Light green

CONCLUSION:

The taxonomical identification of plant material and Pharmacognostical evaluation is important to provide the standards and to avoid adulteration of drugs. Macroscopical and Microscopical characters of the plant used for the identification of the drug. The physicochemical evaluation helps in formulating pharmacopoeial standards, while fluorescence analysis helps in distinguishing the drug in powder form. The physico chemical constants like moisture content, ash values, melting point, extractive values and fluorescence analysis are rarely constant for crude drugs, but they may help in evaluation. Ash value is a criterion to judge the identity and purity of crude drug. The extract obtained by exhausting crude drug is indicative of approximate measure of their chemical constituents. Melting point is one of the parameter to judge the purity of crude drug. To check the moisture content helps prevent degradation.

REFERENCES:

Gamble JS, Flora of Madras, Botanical Survey of India, Calcutta, 1967, 3^rd ed., 954.

Madhava Chetty K, Sivaji K, Tulasi Rao K, Flowering plants of Chittoor District, Andhra Pradesh, India, Students Offset Printers, Tirupathi, 333-334.

Sass JE, Elements of Botanical Micro technique, McGraw Hill Book. Co. New York, 1940, 222.

Johansen DA, Plant Micro technique, McGraw Hill Book. Co. New York, 1940, 523

O’ Brien, T.P., Feder, N. & Mc Cull, M.E. (1964). Polychromatic staining of plant cell walls by toluidine blue- Eds. O. Protoplasma, 59: 364-373.

Esau K, Plant Anatomy, John Wiley & Sons, New York, 1979, 550.

Indian Pharmacopoeia. Vol 2. New Delhi: Controller of Publications; 1996. p. A-53-4, A-89, A-145-9.

Chase CR, Pratt RJ. Fluorescence of powdered vegetable drugs with particular reference to development of system of identification. J Am Pharm Assoc 1949; 38: 32.

Kokoshi J, Kokoshi R, Slama FJ. Fluorescence of powdered vegetable drugs under ultra violet radiation. J Am Pharm Assoc 1958; 47(10): 715.

Ushakumari J, Navas M, Dan M. Pharmacognostic studies on Pellionia heyneana. J Trop Med Plant 2004; 5: 259-61.

Received on 19.12.2013 Modified on 17.01.2014

Res. J. Pharmacognosy & Phytochem. 6(2): April-June 2014; Page 84-87