Pharmacognostical and Preliminary Phytochemical Studies of Nyctanthes arbor – tristis Linn.

V. Suresh¹* and G. Arunachalam ²

¹JKK Muniraja Medical Research Foundation College of Pharmacy, B.Komarapalayam-638 183.

²P.G.P. College of Pharmaceutical Science and Research Institute, Namakkal-637 207.

ABSTRACT:

Nyctanthes Arbor-tristis Linn (Nyctanthaceae) commonly called as night jasmine is used for a wide range of diseases in folk medicine. Only a very limited research has been carried out on the plant, under the present study assumes singular significance and it is supposed to contribute a great deal to the existing literature. The present paper highlights the pharmacognostical studies such as macroscopic, microscopic characters, powder microscopy and fluorescence analysis of bark. It also includes physiochemical constants like ash value, extractive values, loss on drying of the bark powder. These observations would be of immense value in the botanical identification and standardization of the drug in crude form. This study would help distinguish the drug from its other species.

KEYWORDS: Nyctanthes Arbor-tristis Linn, Pharmacognosy, physiochemical constants

INTRODUCTION:

Nyctanthes arbor-tristis Linn (Nyctanthaceae) commonly called as night jasmine, a hardy large shrub or small tree widely distributed in outer Himalayan ranges from Chenab to Nepal, Assam, Burma, Bengal, Central India to Godavari, cultivated in many parts of India. ^1,3,4 It is upto 10m height, young branches have angular stem. Leaves are simple, opposite, ovate, rough and thin. Margins serrate, apex acute. The flowers are delightfully fragrant, sessile, bisexual, hypogynous, corolla tube orange coloured, lobes white, twisted. The seeds are orbicular, thin testa, brown coloured capsule 2cm long and compressed. ^2,5-7 The bark was externally grey or brownish white in colour, rough deep and irregularly fissured. Internally yellowish white or sandal colour, externally granular in texture and internally smooth, 8.2mm thick, bitter taste, odourless, curved or quill in shape, short fracture in outer bark and fibrous fracture in inner bark. ^8,9 The barks are intended for expectorant, anorexia, liver disorder, piles, worm infestation, blood disorder, oliguria, skin diseases fever and snake bite.^1,2,7,10-13 However pharmacognostical information about this plant has not been published, particularly the necessary to define quality control procedures of the Nyctanthes arbor-tristis Linn as raw material. Hence the present investigation deals the pharmacognostical and preliminary phyto chemical evaluation of the N.arbor-tristis Linn. The study includes morphological and anatomical, determination of physico-chemical constants and the preliminary phytochemical evaluation of the different extracts of N.arbor-tristis Linn.¹⁴

MATERIALS AND METHODS:

Collection and Treatment:

The plant was widely cultivated in gardens almost throughout India or the fragrant flowers. For the present work the plant was collected from Sangameswarar temple near Bhavani ( Erode District). The plant was identified by Dr. G. V. S. Murthy, Joint Director of Botanical Survey of India, Southern circle, TNAU Campus, Coimbatore who authenticated the plant from available literature. The bark was collected by felling method, where in trees cut at the base and bark as peeled out. The bark cuttings were collected and washed with water and dried in shade. The bark pieces were powdered by means of wood-grinder and the powder was passed through the sieve No.60 for powder analysis and the coarse fraction was subjected or phytochemical studies.

Chemicals and Instruments:

Compound microscope, Camera lucida, stage and eyepiece micrometer, glass slides, cover slips, watch glass and other common glassware were the basic apparatus and instruments used for the study. Photomicrographs in different magnifications of all necessary cells and tissues were taken with Nikon Lab Phot- 2 microscopic Unit. Some crystals, starch grains and lignified wall photographs were taken under polarized light microscope. Solvents viz. Pet. ether, chloroform, ethanol and reagents viz. Phloroglucinol, Glycerin, HCl, Chloralhydrate and Sodium hydroxide were procured from Ranbaxy Fine Chemicals Ltd., Mumbai, India.

RESULTS AND DISCUSSION:

The powdered bark of Nyctanthes arbor-tristis Linn has been investigated into a systematic way covering pharmacognostical and preliminary phytochemical aspects in an attempt to rationalize its uses drug of therapeutic importance.

Fi: Fissures; Pe: Periderm DR: Dilated ray Scl: Scelerids Cph: Collapsed phloem Neph: Non collapsed phloem.

Pharmacognostical Studies:

Macro and Microscopic Studies:

The surface of the bark is rough, deep, irregularly fissured externally grey or brownish in colour. Internally yellowish white or sandal coloured. Externally granular in texture and internally smooth, odourless and bitter in taste .The fractures are short in outer bark and fibrous in inner bark. The bark has total thickness of 8.2mm and differentiated into outer bark (periderm) and inner bark (secondary phloem).

Table-1: Data for fluorescence analysis of powdered bark of N. arbor-tristis Linn.

Sl. no	Treatment	Day light	UV light (254nm)
1.	Powder as such	Light brown	Light green
2.	Powder + 1N HCL	Yellowish brown	Light green
3.	Powder + Aq. 1N NaOH	Yellow	Dark green
4.	Powder +Alcoholic 1N NaOH	Pale yellow	Yellowish green
5.	Powder + 5% I2 solution	Bluish black	Dark brown
6.	Powder + 50% HNO3	Yellowish orange	Light green
7.	Powder + 50% H2SO4	Yellowish orange	Light green
8.	Powder + Methanol	Pale yellow	Emerald green
9.	Powder + 5% FeCl3 solution	Violet colour	Fluorescent green

Fig 4: Anatomy of collapsed and non-collapsed phloem

Scl: Scelerids; Cph: Collapsed phloem; Ncph:Non- collapsed phloem

Fig 5: TLS view of collapsed phloem

Cph: Collapsed phloem: PhR:Phloem rays; Scl: Scelerids;

Fig 6: TLS view of non-collapsed phloem

STM:Sieve tube members; PhR:Phloem rays; Cph: Collapsed phloem: AP:Axial parenchyma

Fig 7: TLS view of non-collapsed phloem

SP: Simple pits; AP: Axial parenchyma PhR: Phloem rays; STM: Sieve tube members

Fig 8: RLS view of phloem

PhR: Phloem rays Scl: Scelerids PC:Procumbent cells UC: Upright cells

Fig 9.Powder microscopy of bark of Nyctanthes arbor-tristis linn

The periderm is superficial in position, it is wide, measuring 1-2mm thick, at certain places the periderm enters into the inner tissue forming wide bay(Fig.3). The periderm surface is irregularly fissured, the fissures are shallow. The periderm comprises of small, tabular phellem cells which are comprised of dark bands. In places where the phelem is heterogeneous, made up of thin, continuous tangential lines of phelloids (sclereids) and wider, thin walled, squarish or tabular cells (Fig.3.1, 3.2). phelloderm is not evident. Periderm is immediately followed by secondary phloem. The secondary phloem consists of outer collapsed phloem and inner zone of non-collapsed phloem. The collapsed phloem is the widest part of the bark. The region beneath the periderm consists of circular, less compact parenchyma cells, narrow phloem rays and circular, prominent masses of sclereids (Fig 4.1). Further inner to the phloem, here are obliquely radial dark thin lines which represent crushed and obliterated sieve elements or the collapsed phloem (Fig.4.1). The non-collapsed phloem (Fig.4.2) is 450 µm. It consists of intact sieve tube members, axial parenchyma and thin less prominent rays. The sieve tube members are small and are random or radial in orientation. The sieve tube members are 20-30 µm in diameter. The TLS view exhibits the collapsed and non-collapsed phloems.

Table 2: Data for ash values of bark powder of Nyctanthes arbour-tristis Linn

S. No	PARAMETER	%w/w
	ASH VALUES
1.	Total Ash	7.00
2.	Acid Soluble Ash	0.50
3.	Water Insoluble Ash	0.45
4.	Sulphated Ash	3.00

Table: 3 Data for extractive values Nyctanthes arbor-tristis Linn

Sl. No	Extractive value	Percentage (w/w)
1	Alcohol soluble extractive	6.5 %
2	Water soluble extractive	4.76 %
3	Ether Soluble extractive	0.11 %

Table-4: Successive extraction values of various extracts of powdered bark of Nyctanthes arbor-tristis Linn

Sl. No	Extracts	Yield(gm)	% Yield (w/w)
1.	Petroleum ether extract	0.57	0.19%
2.	Chloroform extract	0.46	0.15%
3.	Ethanolic extract	30.0	10%
4.	Aqueous extract	19.0	6.3%

The collapsed phloem shows slightly wider rays which are three to many seriate short and wide, they are homocellular consisting of polygonal compact cells are intact. The sclereids are in thick vertical bands (Fig.5). The sieve tube members are crushed and appear in thick vertical dark lines. The rays are 200-250 µm in breadth. In the non-collapsed phloem region sclereids are absent; the phloem rays are smaller in size. The sieve tube members are short and narrow. They are 250µm in height. The sieve plate is simple and oblique (Fig.6). The phloem parenchyma cells of the phloem have wide simple pits (Fig.7). The RLS view exhibits horizontal ribbon like band of cells. These cells are horizontal oblong or squarish. The rays are homocellular. Some of the rays are heterocellular with horizontal procumbent cells and marginally upright cells (Fig.8). The powdered bark shows phelloid cells, the periderm is broken into small bits in which sclerotic phelloid cells and thin walled phloem cells are evident (Fig.9.1). Calcium oxalate crystals of 2 types are seen in the powder (Fig9.1). sphaerocrytals are spherical bodies formed by many pointed crystals(Fig.9.2). Prismatic crystals include rectangular and spindle shaped crystals, they are equally abundant. (Fig 9.2). The sclerenchyma components of the bark are the stone cell or branchy sclereids. They are squarish, irregular or rectangular in shape. They have thick lignified walls and wide lumen. Numerous simple circular pits are seen in the sclereids (Fig.10).^10-19

Table-5: Fluorescence study of various extracts of powdered bark of Nyctanthes arbor-tristis Linn

Sl. No	Extracts	Day light	UV light(254nm)
1.	Petroleum ether extract	Brown	Light green
2.	Chloroform extract	Yellowish brown	Dark green
3.	Ethanolic extract	Yellowish orange	Light green
4.	Aqueous extract	Brownish black	Dark green

Table: 6: Qualititative phytochemical analysis of various extracts of powdered bark of Nyctanthes arbor-tristis Linn

PHYTOCONSTITUENTS	Petroleum ether extract	Chloroform extract	Ethanol extract	Aqueous extract
Alkaloids	_	+	+	_
Saponins	_	_	_	_
Glycosides	_	_	+	+
Carbohydrates	_	_	+	+
Tannins and phenolic compounds	_	_	+	+
Flavonoids	_	_	+	+
Phytosterols	+	_	_	_
Proteins and amino acids	_	_	_	+
Triterpenoids	_	_	_	_
Fixed oils and fats	+	_	_	_
Gums and mucilage	_	_	+	+

(+): Present, (-): Absent

Fig 10. Powder microscopy of Nyctanthes arbor-tristis linn

Scl: Scelerids; Cr: Crystal; Pi: Pits

Fluorescence Analysis:

The fluorescence studies for the bark powder by treating it with different chemical reagents and the results are presented in Table.No.1. The fluorescence studies for the bark powder by treating it with different solvent extracts of bark were also performed under day light and UV light. The data for ash values are given in Table.No.2. Data for extractive values are given in Table no.3. The above studies enable the identification of the plant material for future investigation and form an important species of drug studies.^20-24

Preliminary Phytochemical Studies:

The shade dried bark of Nyctanthes arbor-tristis Linn was powdered and successfully extracted with different solvents and the data are given in Table.No.4. All the extracts were subjected to fluorescent analysis and the data are given in Table.No.5. The extracts are subjected to preliminary phytochemical test to find out the active constituents and the data are given in Table.No.6. It revealed that presence of alkaloids, carbohydrate, glycosides, phytosterols, fixed oil, tannins, flavonoids, proteins and amino acids, gums and mucilage.^{14.20, 24}.

CONCLUSION:

The plant Nyctanthes arbor-tristis Linn has been examined to gain an insight of its pharmacognostical and preliminary phytochemical nature. The pharmacognostical studies include macroscopical studies, microscopical studies, ash values, extractive values, and loss on drying of crude powdered bark of Nyctanthes arbor-tristis Linn gave valuable information. Especially existances of fluorescent ingredients were identified in fluorescence study of crude powdered bark.

The preliminary phytochemical investigation showed the presence of carbohydrates, glycosides, phytosterols, flavonoids, alkaloids, fixed oil, tannins, proteins, aminoacids, gums and mucilage in various extracts.

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Received on 01.08.2010

Accepted on 11.08.2010

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Research Journal of Pharmacognosy and Phytochemistry. 2(5): Sept.-Oct. 2010, 411-416