INTRODUCTION:

Achyranthes aspera is an erect plant and about 1 m in height. It is common plant as weed and found roadside throughout India. The various vernacular names of the plant are as Sanskrit Aghata, Apamarga, in hindi Chirchira, in marathi Aghada. The various parts like Herb, leaves, seeds and root used in various diseases.Decoction of leaves is a good diuretic found in renal dropsies and general anasarea. The leaf juice is also useful in stomach ache and bowl complaints, piles, boils skin eruptions. In large doses it produces abortion or labour pains¹. It is widely used for asthmatic cough, snakebite, hydrophobia, urinary calculi, rabies, influenza, piles, bronchitis, diarrhea, renal dropsy, gonorrhea and abdominal pain. The seeds are employed as an emetic, purgative, and cathartic, in gonorrhoea, for insect bite and in hydrophobia cough including whooping cough as an anti-asthmatic².

Received on 05.01.2015 Modified on 22.01.2015

Research J. Pharm. and Tech. 8(2): Feb. 2015; Page 212-215

DOI: 10.5958/0974-360X.2015.00039.6

MATERIAL AND METHOD:

Collection and identification of plant material:

In the present study, the leaves of Achyranthes aspera, Linn. (Family-Amaranthaceae), was collected from the local areas of Jalgaon district. The plants were authenticated from Agharkar Research Institute (ARI) Pune. (An autonomous, grant-in-aid research institute of the Department of Science and Technology (DST), Government of India). Soon after authentication, all the crude drugs were dried at room temperature until they free from moisture and subjected to physical evaluation with different parameters.

Macroscopic:

The macroscopic characters such as size, shape, margin, nature, texture, apex, surface, colour, odour, taste were studied for morphological investigation³. The results are reported in Table No. 1

Microscopic:

For microscopy, free hand section leaf was cut and stained according to the prescribed method⁴.

Physicochemical Evaluation:

The extractive values, ash values and loss on drying⁵, fluorescence analysis⁶ were performed according to the official methods. The results are reported in Table No. 2 and 3.

Preparation of extracts:

In the present study, the crude drugs were carefully selected and shade dried. The dried material was reduced to coarse powder in a mechanical grinder and passed through a Sieve No.40 to obtain powder of desired particle size.

About 200 gms of powdered material was subjected to exhaustive extraction successively with petroleum ether, ethyl acetate and ethanol at temperature of 45⁰ – 50⁰C to about 40 cycles per batch for 8 batches. The extraction was continued until the solvent in the thimble became clear then few drops of solvent were collected in a test tube during the completion of the cycle (during syphoning) and chemical test of that solvent was performed. Extraction was completed only when chemical test shows negative results. Finally the drug was be macerated with chloroform water.

After each extraction the solvent was distilled off rotary evaporator and the extract was concentrated at low temperature. These extracts were used for phytochemical investigation.

Phytochemical screening:

The preliminary phytochemical screening of extracts were carried out as per standard procedures^7-13. The results are reported in Table No. 3

RESULT AND DISCUSSION:

The first step towards ensuring quality of starting material is authentication. Thus, in recent years there has been a rapid increase in the standardization of selected medicinal plants of potential therapeutic significance. Despite the modern techniques, identification of plant drugs by pharmacognostic studies is more reliable. The standardization of crude drugs is important before any work carried out. The morphology, microscopy, physicochemical tests are the first step towards establishing the identity and the degree of purity of such materials and should be carried out before any tests are undertaken. The result of this study as follows:

Microscopic characters of leaf :

Transverse section of leaf through the midrib shows a single layered epidermis, on both surfaces; epidermis followed by 4-5 layered collenchyma on upper side and 2-3 layered on lower side. Trichomes of covering and glandular type. Stomata are anomocytic on both sides. Calcium oxalate crystals in rosette form are distributed in ground tissue. Palisade layer and spongy mesophyll can be seen. Ground tissue consisting of thin-walled, parenchymatous cells having a number of vascular bundles; each vascular bundle shows below the xylem vessels, thin layers of cambium, followed by phloem and a pericycle represented by 2-3 layers of thick-walled, non-lignified cells; rosette crystals of calcium oxalate found scattered in ground tissues.

Table No.1: Macroscopic characters of Achyranthes aspera Linn. Leaf.

Sr. No.	Parameters	Observation
	Physical Tests
I	Shape	Elliptic, ovate, rounded sub-acute at the apex, finely and softly pubescent on both sides, entire, petiolate.
II	Size	Thick 3.6-6.8 x 21.3-4.6 cm
III	Colour	Green
IV	Odour	Characteristic
V	Taste	Slightly mucilaginous

Table No.2: Physicochemical analysis of Achyranthes aspera Linn. Leaves.

Sr.no	Parameters	*Achyranthes aspera*
I	Physical tests
	Nature	Coarse powder
	Colour	Green
	Odour	Characteristic
	Taste	Slightly mucilaginous
II	Extractive value
	Pet. ether	3.40 % w/w
	Ethyl acetate	1.42 %w/w
	Alcohol	7.88 %w/w
	Aqueous	8.70 %w/w
III	Loss on drying	7.2 %w/w
IV	Ash values
	Total ash	14.20 %w/w
	Acid insoluble ash	9.10 %w/w
	Water soluble ash	5.10 %w/w

Table No.3: Powder analysis of Achyranthes aspera with different chemical reagents:

Sr. No	Reagent	UV (254nm) Fluorescence	UV (366nm) Fluorescence
1	Powder as such	Light Green	Light Green
2	Powder + 1 N NaOH	Green	Light Green
3	Powder + 50% HCL	Light Green	Green
4	Powder + 50% H₂SO₄	Green	Light Brownish
5	Powder + 50% HNO₃	Green	Dark Brownish
6	Powder + Iodine solution	Dark Green	Dark Brownish
7	Powder + 5% FeCl₃	Yellowish Green	Dark Brownish

Table No.4: Preliminary phytochemical investigation of Achyranthes aspera leaves.

Sr. No

Chemical Tests

PEE

ETAE

ALCE

AQE

Tests for Carbohydrates

Molish's test (General test)

Tests for reducing sugars

a) Fehling's test

b) Benedicts test

Test for Monosaccharides

a) Barfoeds test

Test for Pentose sugars

Tests for Hexose sugars

a) Tollen's phloroglucinol test for galactose

b) Cobalt chloride test

Test for Non-reducing sugars

Tests for Non-reducing polysaccharides (starch)

a) Iodine test

b) Tannic acid test for starch

Test for Gums

Tests for Proteins:

a) Biuret test (General test)

b) Millions test for proteins

c) Xanthoprotein test

d) Test for proteins containing sulphur

e) Precipitation test

Tests for Amino acids:

a) Ninhydrin test (General test)

b) Test for tyrosine

c) Test of tryptophan

d) Test for cysteine

Tests for Fats and Oils:

a) Solubility test

b) Saponification test

c) Paper staining test

Tests for Steroids:

a) Salkowski reaction

b) Liebermann – Burchard reaction

c) Liebermann reaction

Tests for Triterpenoids:

a) Salkowski reaction

b) Liebermann – Burchard test

Tests for Glycosides:

Tests for Cardiac Glycosides:

a) Baljet test

b) Legals test (Test for cardenolides)

c) Test for deoxy sugars (Killer Killani test)

d) Liebermann's test (Test for Bufadenoloids)

Tests for anthraquinone glycosides:

a) Borntraggers test

b) Modified Borntragger's test

Tests for Saponin glycosides

a) Foam test

b) Haemolysis test

Test for Cynogenetic glycosides, Guinard-reaction or Sodium picrate test

Tests for Coumarin glycosides

a) Alkaline reagent test

b) NaOH Soaked paper test

Tests for Flavanoids:

a) Ferric Chloride test

b) Shinoda test

c) Alkaline reagent test

d) Lead acetate test

Tests for Alkaloids:

a) Dragendroff's test

b) Mayers test

c) Hagers test

d) Wagners test

e) Murexide test for purine alkaloids

10)

Tests for Tannins and Phenolic compounds:

a) 5% FeCl₃ solution

b) Lead acetate solution

c) Gelatin solution

d) Bromine water

e) Acetic acid solution

f) Dilute iodine solution

g) Dilute HNO₃

h) Dilute potassium permanganate solution

11)

Tests for Saponins:

a) Foam test

b) Raymonds test

c) Haemolysis test

d) Bromine water test

e) Legal's test

12)

Test for Lipids

+ Present - Absent PEE-Petroleum ether extract, ETAE-Ethyl acetate extract, ALCE-Alcoholic extract, AQE-Aqueous extract.