Pharmacognostic Study of Leaf and Bark of Nerium oleander and Eugenia jambolana

Mohini Upadhye*, Snehal Avhad, Prajkta Dhaygude, S. N. Dhole

Department of Pharmacognosy, P.E. Society’s Modern College of Pharmacy (For Ladies) Borhadewadi, Dehu Alandi road, Moshi, Pune-412 105 Maharashtra

ABSTRACT:

The plant of Nerium oleander is used in various diseases like cancer. It is also used as diaphoretic, diuretic, emetic, expectorant, and resolvant. Eugenia jambolana is used as antibacterial, strengthening of teeth and gums. Both the plants are extensively used as tribal medicines in different parts of country.

As there is no record of pharmacognostical work on such potential drugs, the present work is taken up to produce the pharmacognostical standards. The study deals with the pharmacognostical examinations, morphological, microscopical characters. Phytochemical characteristics of leaves and bark of Nerium oleander and Eugenia jambolana were also determined which include determination of leaf constants, ash values, foaming index, ,swelling index, moisture content and extractive values, foreign organic matter, crude fiber content etc as per official guidelines.

KEY WORDS: Nerium oleander, Eugenia jambolana, bark, leaves, microscopy, physicochemical parameters.

INTRODUCTION:

Nerium oleander (Apocynaceae) is cultivated worldwide as an ornamental plant. All parts of the plant are used as anticancer, cardiotonic, diaphoretic, diuretic, emetic, expectorant, resolvant, skin sternutatory. The leaves of Eugenia jambolana are antibacterial and used for strenghning the teeth and gums. The fruits and seeds are acrid, sour, sweet, tonic, cooling and are used in diabetes. The bark is astringent and is also used in deaction in case of chronic diarrhea and dysentery. It is also used as gargle in sore throat. The fruits are used extensively in the treatment of diabetes. Therefore, the present investigations were planed to study the leaves and bark of Nerium oleander and Eugenia jambolana ^(1).

MATERIAL AND METHODS:

Plant Material:

The leaves and bark of Nerium oleander and Eugenia jambolana were collected from Moshi in the month of October and was authenticated by Dr. Dinesh Shirodkar, Indian Botanical Survey, Yerwada, Pune.

Pharmacognostical Studies^{(2- 4)}

The plants were studies for morphological characters like shape, size, surface, base, margin and venation. The organoleptic characters such as taste, odour and colour of leaves as well barks were studied.

The microscopic studies were carried out using Nikon Camera. The clear free hand sections of leaves and bark which were previously soaked and boiled with chloral hydrate to remove the colouring matter were selected and mounted on a clean glass slide using glycerine and covered with cover slip. The sections were then viewed under low power 10X and 45X. The sections were stained with phloroglucinol and concentrated HCl. It was observed that lignified xylem of vascular bundle were stained pink. The starch grains were stained with dil Iodine. Some of the sections were also stained with Sudan Red for cuticle, Rhuthenium Red for mucilaginous cells and Acetic acid for calcium oxalate.

As a part of quantitative microscopy Stomatal number, Stomatal index, Vein islet number, Vein termination number were determined by using fresh leaves of the plants with the help of camera lucida.

The leaves and bark of Nerium oleander and Eugenia jambolana were powdered and subjected to powder microscopy. Powders were stained with phloroglucinol and concentrated HCl.

Various physicochemical parameters like total ash value, acid insoluble ash value, water soluble ash value, alcohol and water soluble extractive value, moisture content, foaming index, swelling index, foreign organic matter were determined as per the WHO guidelines. The aqueous and ethanol extracts were subjected to preliminary phytochemical screening. ⁽⁵⁾

Table 1 Physical parameters of Nerium oleander and Eugenia jambolana

SR.NO	ANALYTICAL Parameter	Nerium oleander		Eugenia jambolana
		Leaves	Bark	Leaves	Bark
1	ASH VALUE a) Acid insoluble ash b) Water soluble ash	13% w/w 2% w/w	9% w/w 2% w/w	9.25%w/w 13.25%w/w	14.75% 2.75%
2	EXTRACTIVE VALUE a) Water soluble extractive value b) Alcohol soluble extractive value	2% 6%	4% w/w 10% w/w	4%w/w 2%w/w	6% 4%
3	FOREIGN ORGANIC MATTEROF 100 gm	0.5% w/w	2% w/w	1%w/w	2%
4	MOISTURE CONTENT	2% w/w	2% w/w	7%w/w	8%
5	FOAMING INDEX	Less than 100	Less than 100	Less than 100	Less than 100
6	SWELLING INDEX	4	2	3	2

Table 2 Quantitative constants of Nerium oleander and Eugenia jambolana

1	STOMATAL NO.	0.6/mm^2	13/mm^2
2	STOMATAL INDEX	0.9/mm^2	28.8/mm^2
3	VEIN ISLET NO.	12/mm^2	13.4/mm^2
4	VEIN TERMINATION	12/mm^2	29.4/mm^2

Table 3: Preliminary Phytochemical Sreening of Nerium oleander

Sr. No.	Plant Constituents	Nerium oleander Leaf		Nerium oleander Bark
		Aqueous Extract	Alcoholic Extract	Aqueous Extract	Alcoholic Extract
1	Carbohydrates	+	+	+	+
2	Proteins	+	+	+	+
3	Amino acids	+	+	+	+
4	Glycosides	+	-	+	-
5	Fats and oils	+	+	+	+
6	Steroids	+	-	+	+
7	Alkaloids	+	-	-	-
8	Flavonoids	+	-	+	-
9	Phenolic comds and tannins	+	-	-	-

(+) denotes Presence (-) denotes Absence

Table 4: Preliminary Phytochemical Sreening of Eugenia jambolana

Sr. No.	Plant Constituents	Eugenia jambolana Leaf		Eugenia jambolana Bark
		Aqueous Extract	Alcoholic Extract	Aqueous Extract	Alcoholic Extract
1	Carbohydrates	+	+	+	+
2	Proteins	-	-	-	-
3	Amino acids	+	-	-	-
4	Fats and oils	-	+	+	+
5	Steroids	+	-	-	-
6	Volatile oils	+	+	+	+
7	Glycosides	+	-	+	+
8	Tannins	+	-	-	-
9	Flavonoids	+	+	+	+
10	Alkaloids	-	-	-	-

(+) denotes Presence (-) denotes Absence

RESULT AND DISCUSSION:

The leaves of Nerium oleander were 10 to 22 cm long, narrow, untoothed, short stalked, dark grey or green in colour, with prominent midrib, leathery in texture and arise in groups of 3 from the stem. The leaves of Eugenia jambolana were green coloured, elliptical, oblong with acute apex, smooth and shiny texture with characteristic odour and taste.

The bark of Nerium oleander was grayish brown in colour with scars. The smooth longitudinal fractures were observed with bitter odour and characteristic taste. The bark of Eugenia jambolana was found to be reddish brown in colour with silky luster and marked with waving ridges and having bitter taste and characteristic odor.⁽⁶⁾

Transverse section of the leaf of Nerium oleander through midrib showed presence of single layered upper and lower epidermis with anamocytic stomata, uniserrate multicellular covering trichomes and glandular trichomes. Mesophyll showed isobilateral leaf pattern and it was differentiated into palisade and spongy parenchyma. The palisade layer was single layered, elongated and compact cells and the spongy parenchyma is 5 to 8 layered, loosly arranged cells with intercellular spaces. The midrib region showed strips of collenchyma appeared below upper epidermis and above the lower epidermis. A well developed vascular bundle is seen in the midrib region. Xylem was lignified while phloem was unlignified.

The leaf of Eugenia jambolana was isobilateral with covering trichomes and paracytic stomata. Collenchyma was faint brown in colour and present above the lower epidermis. Mesophyll region showed presence of palisade, spongy parenchyma and a well developed vascular bundle was observed in the midrib region.

The bark of Nerium oleander exhibits the typical histological characteristics of the bark. Cork cells were yellowish brown, thin walled followed by phelloderm showing presence of prismatic calcium oxalate crystals and starch grains. The phloem was unlignified and shows sieve tubes, companion cells and parenchyma. Xylem vessels were lignified, thick walled and arranged along with xylem fibers. The medullary rays were radialy arranged from centre to the cortex through the vascular region.

The transverse section of the bark of Eugenia jambolana showed the outer several layers of tangentially elongated cells followed by the phellogen and phelloderm. Continuous band of lignified, pitted, oval cells containing calcium oxalate crystals were present in the cortex region. Medullary rays showed presence of two to three celled thin walled radially elongated parenchymatous cells in the secondary phloem region. ⁽⁷⁾

The powder analysis of Nerium oleander leaves revealed the presence of uniserrate, multicellular covering trichomes, glandular trichomes, lignifies and nonlignified fibers, anamocytic stomata while bark shows dark brownish cork cells, lignified fibers, prismatic calcium oxalate crystals and starch grains.

The powder analysis of leaves of Eugenia jambolana showed the presence of covering trichomes, paracytic stomata, calcium oxalate crystals, xylem vessels, lignified and nonlignified fibers while that of bark showed presence of cork cells. The stone cells were lignified, oval and pitted. Calcium oxalate crystals, medullary rays, lignified and nonlignified fiber were also observed in the powder. ^{(8, 9)}

Results of various physicochemical parameters including ash values, Extractive values, foreign organic matter, moisture content, foaming index, swelling index of leaves and bark of Nerium oleander and Eugenia jambolana are represented in Table no. 1. Results of quantitative microscopical parameters such as stomatal number, stomatal index, vein islet number, vein termination number are represented in Table No. 2. Table No. 3 and 4 represent the results of preliminary phytochemical screening of aqueous and alcoholic extracts of leaves and bark of Nerium oleander and Eugenia jambolana.

Thus, macro and micro morphological standards discussed for Nerium oleander and Eugenia jambolana can be considered as distinguishing parameters to identify the authenticity of drugs in herbal industry or trade and thus can be included as microscopic standards in Indian Herbal Pharmacopoeia.

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Received on 20.08.2012

Modified on 02.09.2012

Accepted on 18.09.2012

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 4(5): September – October 2012, 277-279