INTRODUCTION:

Zanthoxylum tetraspermum wight and arn(rutaceae) is a thorny stout, aromatic, climbing shrub, with brown bark having short recurved prickles, found in the western ghats in the Nilgiri, Annaimalai hills and in Kollihills (Namakkal District, Tamilnadu) at altitudes of 1,200-1,800 m, also habitat in Kerala and Karnataka. The wood is yellowish white and soft. The plant is credited in Srilanka with stimulant, astringent and digestive

properties and is prescribed in dyspepsis and diarrhea.^1-3 8-acetonyldihydronitidine, 8-acetonyldihydroavicine, liriodenine, sesamin, lichexanthone, (+)-piperitol-gamma–gamma-dimethylallayl ether have been reported.⁴ From the literature survey there is no evidence for the pharmacognostical studies of this plant for easy identification. Therefore the present investigation was planned to study the pharmacognostical aspects of zanthoxylum teraspermum wight and arn.

MATERIALS AND METHODS:

PLANT MATERIAL

Zanthoxylum tetrasperm wight and arn specimens for the proposed study were collected from Cholakkadu village of Kollihills in Namakkal district on August 2007 and positively identified and authenticated by G. V. S. Murthy, Joint Director, Botanical Survey of India, TNAU southern circle, Coimbatore (No. BSI/SC/5/23/07-08/Tech 715). A herbarium specimen of the plant was preserved in the department of pharmacognosy of our institute for further reference.

Care was taken to select healthy plants and for normal organs. The required sample of different organs were cut and removed from the plant and fixed in FAA (Formalin 5 ml + acetic acid 5 ml + 70% ethyl alcohol 90ml). After 24 hrs of fixing the specimens were dehydrated with graded series of tertiary -butyl alcohol.⁵ Infiltration of the specimens was carried by gradual addition of paraffin wax (melting point 58-60°C) until TBA solution attained super saturation. The specimens were cast into paraffin blocks.

PHARMACOGNOSTICAL STUDIES:

The paraffin embedded specimens were defined with the help of rotatory microtome. The thickness of the sections was 10-12µm. Dewaxing of the sections was by customary procedure.⁶ the sections were stained with toluidine blue.⁷ since toluidine blue is a polychromatic stain, the staining results were remarkably good and some phytochemical reactions were also obtained. The dye rendered 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, whenever necessary sections were also stained with safranin and fast-green and IKI (for starch). For studying the stomatal morphology, venation pattern and trichomes distribution, paradermal sections as well as clearing of leaf stained with 5% sodium hydroxide or epidermal peeling by partial maceration employing Jeffrey’s maceration fluid were all prepared (sass, 1940) Glycerin mounted temporary preparations were made for macerated or cleared powdered materials of different parts were cleared with NaOH and mounted in glycerin medium after staining. Different cell component were studied and measured.

Fig.1 T.S of leaf Zanthoxlum tetraspemum wight and arn through midrib with lamina

Fig.2 T.S of leaf lamina of Zanthoxylum tetraspemum wight and arn through lateral vein

Fig.3 T.S of leaf lamina of Zanthoxylum tetraspemum wight and arn enlarged

Fig.4 T.S of leaf midrib of Zanthoxylum tetraspemum wight and arn

Table1: Analytical parameters of Zanthoxylum tetraspermum wight and arn

PARAMETER	RESULT
Stomatal number	Upper epidermis: absence Lower epidermis: 4-6
Stomatal index	Upper epidermis: absence Lower epidermis: 15-78%
Vein islet number Vein termination number Total ash Water soluble ash Acid soluble ash Sulphated ash Water soluble extractive value Alcohol soluble extractive value Foaming index	3 17 13.5 % w/w 1.5 % w/w 2.0 % w/w 17.0 % w/w 11.5 %w/w 9.0 %w/v 150

RESULTS AND DISCUSSION:

LEAF:

In cross sectional views the leaflet exhibits distinct midrib and uniformly thick lamina, the midrib has an adaxial widely conical lump and a semicircular abaxial part (Fig 1). The midrib in 800µm thick in vertical and the abaxial part in 750µm wide, the lamina is 200 µm thick.

LAMINA:

The lamina (Fig 2) has prominent and adaxial and abaxial epidermal layers. The adaxial epidermis in 40 m.m thick the cells are semicircular with flat outer tangential walls, the cuticle is prominent. The leaf axial epidermis is slightly narrow and is 20-30µm thick. The cells are rectangular or circular. Some of the cells have divided layers transverse wall. So that the epidermis become two layered. The mesophyll tissue is differentiated into adaxial palisade zone and abaxial spongy mesophyll zone. The palisade zone is 50µm in height; the cells are narrowly cylindrical and loosely arranged. The spongy parenchyma zone has four (or) five layers of lobed loosely arranged cells forming wide air-chambers, wide, circular. Secretory cavities are often seen in the spongy mesophyll tissue. The cavities do not have special secretory epithelial cells (Fig 3). The cavity is 20 µm wide.

MIDRIB:

Midrib has large, semicircular, papillate epidermal cells which are densely filled with dark tannin content (Fig 4). The adaxial epidermis is 20µm thick and the abaxial layer is 30-40 µm thick. The adaxial portion consists of about five layers of thick walled compact cells. The cell layers become reduced to a single hypodermal layer cell the lateral portion of the midrib. In abaxial part also there are about three layers of compact collenchymas cells. Inner to the collenchymas portion are two (or) three layers of large circular parenchyma cells. In between these two zones there is a thick dark are of crushed line of cells. The vascular system consist of an abaxial wide, bowl shaped structure and adaxial thick plate. Xylem elements occur in close, radially parallel rows inter mixed with fibers. The xylem elements are thick walled, lignified and angular in outline (Fig 4). Phloem occurs in fairly wide band on the outer part of the xylem. External to the abaxial bowl and adaxial plate of vascular tissues, there is a thick selerenchyma band, which is a continuous (or) discontinuous mass.

Microscopic study of Zanthoxylum tetraspermum leaf powder showed the presence of unicellular, lignified trichomes, anisocytic stomata, simple starch grains and prismatic type of calcium oxalate crystals. The stomatal values, vein islet number, vein termination number, total ash, water soluble ash, acid insoluble ash, sulphated ash, water and alcohol soluble extractive values and foaming index of leaves powder are given in Table-1. The qualitative chemical test performed revealed the presence of alkaloids, glycosides, sterols, triterpenoids, saponins, fixed oils and fats.

CONCLUSION:

The present study on pharmacognistical characters of zanthoxylum tetraspermum wight and arn might be useful to supplement information in regards to its identification. As there is no pharmacognostical work reported for this plant, this study will reveal identity, purity and quality of the plant material.

REFERENCES:

1. Ambasta, The Wealth of India, Rawmaterials, Vol-xI, CSIR, New Delhi,1998,pp.17-25.

2. Chaudha, Y.R., The Useful Plants of India, Publication and Information Directorate, CSIR, New Delhi, 1994, pp.697-699.

3. Hajra,P.K., Nair,V.J and Daniel,P., Flora of India, Vol-4,Botanical Survey of India, BSI, Calcutta,1997,pp.389-390.

4. Nissanka,A.P.K., Karuharatne,V., Rathnayke Bandara,B.M., Phytochemistry, Vol-56(8),2001,.

5. Sass,J.E., Elements of Botanical Microtechnique, Mc Graw Hill Book Co, New York,1940,pp.222.

6. Johansen,D.A., Plant Microtechnique, Ms Graw Hill Book Co, New York,1940,pp.523.

7. O’Brien,T.P., Feder,N. and Mc Cull,M.E., Polychromatic Staining of Plant Cell Walls by Toluidine blue-o,Protoplasma,1964,pp.373.

Received on 17.07.2009 Modified on 20.09.2009

Research J. Pharm. and Tech. 3(1): Jan.-Mar. 2010; Page 106-108