REVIEW ARTICLE ON ANTIMICROBIAL, ANXIOLYTIC AND MUSCLE RELAXANT ACTIVITY OF ECLIPTA ALBA (BHRINGRAJ)

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PHARMACOLOGICAL PROPERTIES:
Crude extract:

The crude extract has been found to have wound healing properties. It been reported to counteract CCl4-induced inhibition of the hepatic microsomal drug metabolizing enzymes. The loss of hepatic lysosomal acid phosphatase and alkaline phosphatase by CCl4 was significantly restored by Eclipta alba. The study shows that hepatoprotective activity of Eclipta alba is by regulating the levels of hepatic microsomal drug metabolizing enzymes. The fresh plant is used as self medication by AIDS patients in southern Thailand and showed potential as a therapeutic agent against Giardia intestinalis infections. The leaf extract showed hypolipedemic activity in atherogenic diet induced hyperlipedemic rats. It has antimicrobial and antioxidant properties. 3% extract of Eclipta alba is used in pilex formulation with other ingredients. It has been reported to decrease bleeding time. Leaf extract has been used in edema. It is used in the treatment of paronychia.

Pharmacological activities of the chemical constituents:

Sl.No

  Chemical  constituents

  Pharmacological activites


1

Wedelolactone

Antihepatotoxic Antibacterial, Trypsin Inhibitor, Antivenom

2

Eclalbosaponins

Hair revitalizing,

Antiproleferative, Antigiardial

3

Demethylwedelolactone

Antihepatotoxic, Antihaemorrhage, Antivenom, Dye (cosmetic)

4

Dasyscyphin C

Antiviral,Anticancer

5

Eclalbatin

Antioxidant

6

Ecliptalbine, verazine

Lipid lowering, Analgesic

Other pharmacological activities:
It has been reported that the importance of free carboxylic acid at C-28 position in echinocystic acid derivatives from the methanolic extract Eclipta prostrata showed antifibrotic activity. Ethanolic and ethyl acetate fractions of Eclipta prostrata were tested for its antibacterial activities against Escherichia coli, Klebsiella pneumoniae, Shigella dysenteriae, Salmonella typhi, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. Eclipta prostrata is combined with a non-plant material which is used to bath children suffering from malnutrition for 9 days and used as self medication by AIDS patients in southern Thailand. 16 parts of Eclipta prostrata (bhringaraj), 1 part of Triphala formula {Emblica officinalis (amalaki), Terminalia chebula, (haritaki), Terminalia belerica (bibhitaki)}, 1 part of Caltropis gigantean (arka) and 1 part of Smilax officinalis (sariva) mixed with 80 parts of sesame oil and boiled to make a medicated oil which is reported to be used in skin diseases.

Combination therapy:
Eclipta alba (whole plant), Mimosa pudica (whole plant), Vitex negundo (whole plant), and Solanum nigrum (aerial parts) possessed styptic and anti-inflammatory properties and help in regeneration of the vascular endothelium36. Combination of Herbs like Anethum sowa (Shatapushpa), Piper longum (Pippali mool), Valeriana wallichii (Tagar), Cassia fistula (Aragvadh), Withania somnifera (Ashwagandha) and Triphala (A herbal combination of three fruits) with Eclipta alba (Bhringaraj) pacify the aggravated Vata dosha and combination with Elaeocarpus ganitrus (Rudraksha), Herpestris monniera (Brahmi) showed a tranquilizer effect

LITERATURE REVIE
1.David Banji et.al reported the Impact of aqueous extract of Eclipta alba on material aggression in rats
2.Anal K Jha, Kamlesh Prasad, et.al studied the biosynthesis of silver nano particles using eclipta leaves.
3.R.K.Roy,Mayank thakur et.al revealed the hair growth promoting activity of Eclipta alba in male albino rats.
4.Otilia J.F. Lobo David Banji et al evaluated the antiaggressive activity of Eclipta Alba in experimental animals.
5.Saroj Bapna, et.al.revealed Anti malerial activity of Eclipta alb AGAINST PLASMODIUM BERGHEI.
6.Vasavi Rangineni et.alreveiled diuretic,hypotensive and hypocholesterolemic effects of Eclipta Alba in mild hypertensive subjects :a pilot study.
7.V.D.Thakur and S.A.Mengi reveiled the Neuropharmacological profile of eclipta alba (linn.) Hassk.

MATERIAL AND METHOD:
COLLECTION OF BHRINGRAJ PLANT
.The proper source of the crude drug was identified and the bhringraj plants were collected from Jabalpur division of M.P. State.during JULY2011.

HARVESTING:. the bhringraj plants were collected and washed properly and other plants like grass etc. Were removed from it.

DRYING: Drying consists of removal of excess moisture content of crude drug,so as to improve its quality and make it resistant to microbial contamination.Hence the collected plants were dried for sufficient amount of time under shade to prevent loss of volatile constituents.

PREPARATION OF EXTRACT: The mature leaves were cleaned, dried under shade and were ground into a coarse powder. 50 gram of the powdered plant was successively extracted with ethanol and water by hot soxhlet  method. The extraction was carried out for 4 hours for each solvent. The ethanolic as well as aquous extract were concentrated to get the dry residue.The dried residue were weighed.The extracts were condensed using rotary vaccum evaporator followed by vaccum evaporator and stored in a desiccators.EXTRACTIVE VALUE

EXTRACT

WEIGHT OF HERB

EXTRACTIVE VALUE

Ethanolic extract

50gm

6.328%

Aquous extract

50gm

4.677%

4.3.2  PRELIMNARY PHYTOCHEMICAL INVESTIGATION:
Identification Of The Plant Constituents By Phytochemical Tests:
Ethanolic extract is subjected to various preliminary phytochemical analysis to test for the presence or absence of various phytoconstituents by the following tests.

1. Test for alkaloids: To the extract dilute hydrochloric acid will be added and filtered. The filtrate will be treated with various alkaloid reagents.
a) Mayer’s test:
The filtrate will be treated with Mayer’s reagent: appearance of cream colour indicates the presence of alkaloids.
b) Dragendroff’s test:
The filtrate will be treated with Dragendroffs reagent: appearance of reddish brown precipitate indicates the presence of alkaloids.
c) Hager’s test:
The filtrate when treated with Hager’s reagent, appearance of yellow colour precipitate indicates the presence of alkaloids.

2) Test for carbohydrates and reducing sugar:  The small quantities of the filtrate will be dissolved in 4ml of distilled water and filtered. The filtrate will be subjected to
a) Molisch’s test: 
A small portion of the filtrate will be treated with Molisch’s reagent and sulphuric acid. Formation of a violet ring indicates the presence of carbohydrates.
b) Fehling’s test: 
The extract will be treated with Fehling’s reagent A and B. The appearance of reddish brown colour precipitate indicates the presence of reducing sugar.
c) Benedict’s test: 
The extract will be treated with Benedict’s reagent; appearance of reddish orange colour precipitate indicates the presence of reducing sugar.
d) Barfoed’s test: 
The extract will be treated with barfoed’s reagent and heated. Appearance of reddish orange colour precipitate indicates the presence of non reducing sugars.

3) Test for proteins:
a) Biuret test: 
The extract will be treated with copper sulphate solution, followed by addition of sodium hydroxide solution; appearance of violet colour indicates the presence of proteins.
b) Millon’s test: 
The extract will be treated with Millon’s reagent; appearance of pink colour indicates the presence of proteins.

4) Test for tannins:  The extract will be treated with 10% lead acetate solution; appearance of white precipitate indicates the presence of tannins.

5) Test for phenolic compounds:
a) The extract will be treated with neutral ferric chloride solution; appearance of violet colour indicates the presence of phenolic compounds.
b) The extract will be treated with 10% sodium chloride solution; appearance of cream colour indicates the presence of phenolic compounds.

6) Test for flavonoids:
a) 5ml of extract will be hydrolyzed with 10%sulphuric acid and cooled. Then, it will be extracting with diethyl ether and divided in to three portions in three separate test tubes. 1ml of diluted sodium carbonate, 1ml of 0.1N sodium hydroxide, and 1ml of strong ammonia solution will be added to the first, second and third test tubes respectively. In each test tube. Development of yellow colour demonstrated the presence of flavonoids.
b)Shinoda’s test: The extract will be dissolved in alcohol, to which few magnesium turnings will beaded followed by concentrated HCL drop wise and heated, and appearance of magenta colour shows the presence of flavonoids.

7) Test for glycosides :  When a pinch the extract was treated with glacial acetic acid and few drops of ferric chloride solution, followed by the addition of conc. Sulphuric acid, formation of ring at the junction of two liquids indicates the presence of glycosides.

8) Test for saponins
Foam test : 
About 1 ml of the extract was diluted to 20 ml of with distilled water and shaken well in a test tube. The formation of foam in the upper part of test tube indicates the presence of saponins.

PREPARATION OF STOCK SOLUTION:
Stock solution was prepared  by dissolving 100mg of extract in 100ml of water.

SELECTION OF DOSE:
Dose was selected according to LD 50 of Bhringraj.LD 50 was found out to be 3000mg/kg.
1st dose was selected as 1/10th of LD 50-300mg/kg.
2nd dose was selected as ½ of LD 50-150mg/kg.

TESTS
Individual tests were performed for
1.Anti microbial,
2.Antianxity &
3.Muscle relaxant activity.

ANTI MICROBIAL ACTIVITY

Two procedures are generally employed in microbial assay

1.CYLINDER PLATE METHOD(CUP):  This is based on measurement of the diameter of microbial growth inhibition surrounding the cylinders containing various dilutions of the test compound which are placed on the surface of a solid nutrient medium previously inoculated with a culture of suitable microbe.Inhibition produced by the test compound is compared with that produced by known concentration of a refrence standard.

TURBIDIMETRIC METHOD:  Based on inhibition of microbial growth as in indicated by measurement of turbidity(transmittance)of suspension of a suitable micro-organism in a fluid medium,to which have been added graded amounts of the test compounds.Changes in the transmittance produced by the tested compounds are compared with those produced by known concentration  of standard.


Anti-microbial activity of bhringraj:

Requirements: Aquous as well as ethanolic extract of eclipta alba, B.subtilis, E.coli.nutrient agar medium,cup & plate.

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