Professional Documents
Culture Documents
Materials and Methods
Materials and Methods
Plant:
The whole plants of Erythrina indica were collected from country drug and herbal shop. The plant
was authenticated by plant anatomy research centre, chennai, Tamil Nadu, India.
Preparation of the plant extract:
The whole plants of Erythrina indica were shade dried and coarsely powdered and was
extracted by using aqueous as a solvent in a soxhlet apparatus. The solvent was completely removed by
vaccum and semisolid mass was obtained (11% w/w with respect to the powdered material) the extract
was dried under reduced pressure using rotary flash evaporator and stored in refrigerator for further
studies.
Preliminary Phytochemical Screening:
100mg of aqueous extract was dissolved in distilled water and the following test was done
separately.
TEST FOR ALKALOIDS:
Mayer’s Test:
Reagent:
Solution A: 1.358g of mercuric chloride was dissolved in 60ml of distilled water.
Solution B: 5g of potassium iodide was dissolved in 10ml of distilled water.
Solution A and B were mixed and made up to 100ml with distilled water. 0.5ml of Mayer’s
reagent (potassium mercuric iodide) was added to the filtrate of Erythrina indica. The presence of
alkaloids was confirmed by the formation of cream precipitate.
INVITRO STUDIES:
Free Radical Scavenging activity of Erythrina indica:
DPPH assay:
The free radical quenching capacity of aqueous extract of Erythrina indica was
determined by the method involving the bleaching of stable DPPH (Ursini et al, 1994). A reaction
mixture containing methanol, DPPH (10Mm, 30µl) and 100µl of various concentrations of Erythrina
indica (0.01, 0.1 and 1.0mg in dimethyl sulfoxide, DMSO) was allowed to stand at room temperature
for 30mins before mixing with redistilled distilled water (1ml) and toluene (3ml). The solution was
then centrifuged, and the absorbance of the upper phase was read at 517nm against a blank without
Erythrina indica prepared and processed as described above. Vitamin C served as standard and
compared with the extract. The free radical scavenging activity was calculated as given below.
buffer, before being incubated at 25∘C for 150 mins. The amount of NO- produced by
sodium nitro prusside was measured by Griess reagent (1%) (Sulphanilamide in
5% phosphoric acid and 0.1% N- 15- naphtyl ethylenediamine dihydrochloride in
distilled water). 0.5ml of the incubation mixture was mixed with same volume of
Griess reagent. The absorbance of the mixture was read at 550nm. Vitamin C
served as standard and compared with the extract. The percentage of inhibition
per each concentration of the extract was calculated as given below.