Pharmacognostical and Preliminary Phytochemical Studies of Argyreia nervosa Burm

Ethnobotanical Leaflets 13: 293-300. 2009.

Pharmacognostical and Preliminary Phytochemical Studies of Argyreia nervosa Burm.

Krishnaveni, A¹ and Santh Rani Thaakur²

¹Department of Pharmacognosy, College of Pharmacy,

Madurai Medical College, Madurai,Tamilnadu, India

²Deparment of Pharmacology, Sri Padmavathi Mahila Visvavidyalayam,

(Women’s University), Tirupati, Andhra Pradesh, India

¹For correspondence, E-mail: [email protected] com

Issued 15 February 2009

ABSTRACT

Pharmacognostical parameters for the leaves of Argyreia nervosa Burm were studied with the aim of drawing the pharmacopoeial standards for this species. Macroscopical and microscopical characters, physio-chemical constants, quantitative microscopy parameters, extractive values with different solvents, fluorescence analysis of dry powder, its reaction after treatment with chemical reagents under visible light and UVlight at 254 nm and 366 nm. Preliminary phyto chemical screening on the leaves Argyreia nervosa Burm were studied. The determination of these characters will help future researchers in their Phytochemical as well as Pharmacological analyses of this species.

Keywords: Argyreia nervosa, Convolvulaceae, pharmacognostic, phytochemical.

INTRODUCTION

Argyreia nervosa Burm is a climbing shrub with woody tomentose stem,found mainly in Deccan,Karntaka and East slopes of the West Ghats at an altitude of 900m¹. It is commonly known as Elephant creeper and in Samudra- sok Hindi².Traditinally leaves used by Rajasthani tribes to prevent conception³ . Seeds of Argyreia nervosa found to possess hypotension,spamolytic ⁴, and anti-inflammatory activity⁵. Chemical analysis revealed the presence of triterpenoids,flavanoids,steroids and lipids⁶. Roots of Argyreia nervosa proved the immunomodulatory activity against the myelosuppressive effects induced by Cyclophosphamide⁷. 24R-ergost-5- en-11-oxo-3 beta-ol alpha –D glucopyranoside xylose was isolated from seeds of Argyreia nervosa known as Argyreioside ⁸.

Table 1. Ethnomedical information on Argyreia nervosa Burm⁹.

Parts	Uses
Root	Appetitiser,anaemia,aphrodisiac,anti–inflammatory,brain-tonic,cardiotonic,cerebral disorders, diabetes,expectorant,obesity,syphilius, tuberculosis , ulcers and wounds.
Stem bark	No report available
Fruit	No report available
Seed	No report available
Leaves	Prevents conception, antiphlogistic
Flowers	No report available

But no pharmacognostical work has been done so far. Therefore, an attempt has been made to study the Pharmacognostic parameters on the leaves of Argyreia nervosa Burm in both whole form and powdered form.

Table 2. Macroscopy of Argyreia nervosa Burm.

Parts	Observation
Bark	Woody tomentose
Flowers	pink
Fruit	Fresh /globular
Seed	Black
Leaves	15-22 cm.
Appearance	Silky & tomentose /glabrous beneath
Shape	Cordate
Length/height	7. 5 -18 by 2.5-10 cm.
Apex/base	Acute /cordate.
Petiole	petioled
Surface	Glabrous
Arrangement	alternate
Venation	Reticulately Pinnate

Materials and Methods

Plant material

The plant material was collected from the foothills of Tirumlahills, Tiruapthi.A.P.in the month June2007.The plant was identified and authenticated by Dr.Madhav Shetty,Taxnomist,Dept of Botany, S.V.University, Tiruapthi. A herbarium was preserved in the department for further reference. The leaves were separated, dried, coarsely powdered passed through sieve no 40 and stored in a closed container for further use. All reagents used were of analytical grade obtained from S.D. Fine Chemicals Ltd., Mumbai.

Methods

The macroscopical characters (size, shape colour, odour, texture, venation margin, base, apex and petiole) of the leaves were observed¹⁰. Then, anatomical study, powder was identified with routine reagents to study the lignified cells, trichomes, stomata, fibres etc. Quantitative microscopy was determined by methods prescribed by Trease and Evans¹¹.

The ash values, extractive values with various reagents and were determined as per the Indian Pharmacopoeia¹². The behaviour of powdered leaves with various chemical reagents was studied ^13,14. The fluoroscence characters of the powder with various acids were observed under visible light and UV light as per the proceduere^15. Measurement of vein islet number, vein termination number, stomatal number, stomatal index and length of trichome were determined. Extractive values were performed with various solvents like petroleum ether, chloroform, ethyl acetate, alcohol and water was performed as per Ayurvedic Pharmacopoeia¹⁶. Prelimnary phytochemical tests of the powder/extracts were performed using specific reagents through standard procedures¹⁷.

Results

Analysis and Discussion

Colour -green, odour - odourless, taste – slightly bitter , size 15-22cm in length, cordate in shape, tomentose-surface, acute-apex, base-cordate, entire-margin, paripinnate venation and petioled . The physical constants such as total ash value

(4.3 %) acid insoluble ash (1.6%) water soluble ash (3.94%) which are specific identification for this species. The soluble extractive values respectively, which indicates the nature of constituents present. Quantitative microscopical study also give valuable information regarding specific leaf constants such as vein islet(10.2/mm²),vein termination number(12.6/mm²⁾ stomatal number(4.5/mm² and 16 /mm²) upper and lower epidermis respectively. Length of trichome(12.98µ--59.38 µ--101.9µ ) The behaviour of leaf powder upon treatment with different chemical reagents was also observed and reported in table no 5. Fluoroscence studies of powder with various reagents revealed the presence of green & orange fluoroscence with Conc. sulphuric acid and glacial acetic acid respectively under UV light at 254 nm and 366 nm. The powder microscopy revealed the presence of glandular &covering trichomes, xylem fibres, epidermal cells, cork cells, vessels with bordered pits,,xylem vessels with spiral thickenings were recorded.

Powder analysis of Argyreia nervosa Burm.

It is pale green, fine, odourless powder with slight bitter taste. The powder microscopy revealed the presence of glandular &covering trichomes, xylem fibres, epidermal cells, cork cells, vessels with bordered pits,xylem vessels with spiral thickenings were recorded. The various qualitative chemical tests (table no:7 ) have shown the presence of triterpenoids, saponins, sterols, flavanoids, carbohydrates phenols, tannins and in large amount whereas aromatic acids, , gums and mucilage and volatile oils were totally absent in the leaf extract of this plant.

Table 3. Determination of Ash Values of Argyreia nervosa Burm.

S. No.	Ash type	Percentage of Ash
1.	Total ash	4.3% w/w
2.	Acid insoluble ash	1.6% w/w
3.	Water soluble ash	3.94% w/w

Table 4. Determination of Extractive Values of Argyreia nervosa Burm.

S. No.	Solvent	Percentage of extractive
1.	Petroleum ether	3.16% w/w
2.	Chlororform	0.8% w/w
3.	Ethyl acetate	1.4% w/w
4.	Ethanol	0.2% w/w
5.	Water	7.6% w/w

Table 5. Determination of phyto constants of Argyreia nervosa Burm.

Leaf constants	Report
Vein islet number	10.2/mm²
Vein termination number	12.6/mm²
Stomatal index (upper epidermis)	4.5/mm²
Stomatal index (lower epidermis)	16/mm²

Similarly the fluroscence characterstic of the leaf powdered leaf, when treated with various chemical reagents and its extracts have also been extensively studied.The extractive values of the powder with different solvent was determined and its result was reported in table no: The various qualitative chemical tests (table no ) have shown the presence of sterols, flavanoids, phenols, tannins and saponins in large amount whereas aromatic acids, carbohydrates,triterpenoids gums and mucilage and volatile oils were totally absent in the leaf extract of this plant.

Powder as such:

Colour : Dark green.

Taste: Slightly bitter.

Odour: Characteristic .

Table 6. Behavioural characterstics of powdered leaves of Argyreia nervosa with different chemical reagents.

S. No.	Particulars		Under Visible light	U.V. light
				Short wavelength		Long wavelength
1.	Powder as such		Dull green	Dark green		-----
2.	Powdered drug + Conc. HCl		Dull green	Pale green		-----
3.	Powdered drug + Conc. H₂SO₄		Dull green	Pale green		Green
4.	Powdered drug+ Conc. HNO₃		Yellow	Dull green		-----
5.	Powdered drug+ Glacial Acetic acid		Dull green	Pale green		Orange
6.	Powdered drug+ Aqueous NaOH		Dark green	Dark green		-----
7.	Powdered drug + NaOH (Alcoholic)		Dark green	Dark green		-----
8	Powdered drug + 10%Hcl		Dull green	Dull brown		-----
9.	Powdered drug + 10% H₂SO₄		Dull brown	-----		-----
10.		Powdered drug + 10% HNO₃	Dull green		Dark green	-----
11.		Powdered drug + 10% Glacial Acetic acid	Dark green		Dark green	-----
12.		Powdered drug + Ferric chloride(Aqueous)	Dark green		Dark green	-----
13.		Powdered drug + Ferric chloride(Alcoholic)	Dark green		Dark green	-----

Table 7: Preliminary phytochemical screening of Argyreia nervosa Burm.

S. No.	Tests	Powder + Water	Ethanol extract	Water extract
1.	Alkaloids:
	Dragendroff’s test Mayer’s test Hager’s test Wagner’s test	+ ve + ve + ve + ve	+ ve + ve + ve + ve	+ ve + ve + ve + ve
2.	Carbohydrates: Fehling’s test Molish test	+ ve + ve	+ ve + ve	+ ve + ve
3.	Gums/Mucilage
	Water Alcohol	-ve -ve	- ve - ve	- ve - ve
4.	Tannins:
	Aq. FeCl₃ Test Alc. FeCl₃ Test	+ ve + ve	+ ve + ve	+ ve + ve
5.	Flavonoids:
	Lead acetate test Shinoda test Mg/Hcl	+ ve + ve + ve	- ve - ve - ve	+ ve +ve + ve
6.	Saponins:
	Foam Test Lead acetate test	+ ve + ve	+ ve + ve	- ve + ve
7.	Sterols:
	Salowaski test Libberman Burchad test	+ ve + ve	+ ve + ve	+ ve + ve

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Tests

Powder + Water

Alkaloids: