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Int. J. of Marine Science 2012, Vol.2, No.6, 43
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50
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47
Table 4 Screening of selected sponges for chemical constituents
Sample
no.
Test
Reaction
Present(+)/absent (
-
)
1
Test for Steroids:
Cold extract + minimum amount of CHCl
3
+ 3
drops of acetic anhydride + 2 drops of Conc.
sulphuric acid
Purple colour solution changing to blue
or green
+
2
Test for Triterpenoids:
Cold extract + piece of tin + 3 Drops of thionyl
chloride
Violet or purple colour solution
+
3
a
b
c
Test for Reducing sugar:
Cold extract + equal volume of Fehling A&B +
heated over water bath
Cold extract + Tollens reagent and heated over bath
Cold extract + Molish reagent
Red precipitate
Silver mirror purple colour
Purple colour
+
4
a.
b.
Test for Aalkaloids:
Cold extract + 2N HCl (aqueous layer decanted)
+ 2 drops of Mayers reagent (KI + HgCl)
Cold extract + acetic acid (aqueous layer
decanted) + few drops of Dragendroff’s reagent
Pale yellow or white precipitate
Orange or red orange precipitate
+
5
Test for Phenolic compound:
Cold extract + Neutral FeCl
3
An intense blue or violet coloration
+
6
Test for Saponin:
Cold extract + water mixture shaken vigorously
Formation of foamy layer
+
7
Test for Xantho proteins:
Cold extract + concentrated Nitric acid + excess
Ammonia
Red orange precipitate
+
8
Test for Tannins:
Cold extract + basic lead acetate
White precipitate
+
9
Test for Flavanoids:
Cold extract + bit of Mg
2+
drops of Con. HCl
Heated and then cool
Red or orange red color
+
10
Test for Aromatic acids:
Cold extract + Saturated Sodium bicarbonate
Brisk effervescence
+
3 Materials and Methods
3.1 Collection of sponges, sample preparation and
extraction
The sponges 22 species were collected from the low
inter tidal pools at Bay of Bengal from Gulf of
Mannar Biosphere reserve of Tuticorin coast (8° 47
′
N, 78° 8
′
E) and Rameswaram coast (9° 28
′
N, 79°
12
′
E) at 4~5 m depth from Tamil Nadu, India. The
sponges were collected by an eco-friendly bulk
collection by catch in the fishing nets. From the all
twelve sponges identified, tissues samples were
incised out and (100 g) were washed with sea water,
air dried and chopped into small size before being
ground into fine paste. Using the paste the ethyl
acetate (EtOAc), and methanol (MeOH) were carried
out. The extraction was carried out in triplicates for 48
h. the extract was stored in dark container and left in
deep freezer at
－
20
℃
. After 48 h the extract was