reagent and glucose was estimated as described
earlier. The amount of cellulose was calculated from
the standard curve and expressed in mg cellulose g-1
dry wt.
Hemicellulose content was estimated according
to the procedure of Loomis and Shull (1937). One g
of dried powder was extracted at least twice in 20
ml of hot 80 % ethanol for 10 min. The supernatant
was discarded each time; the residue left was dried
at 70° C in an oven. The dried sugar free residue
hydrolyzed with (1N) HCl in a 25 ml open conical
flask for 1 h at 15 LbsPsi in an autoclave. The
reducing sugar liberated was determined using
anthrone reagent. The amount of sugar multiplied
by 0.9 to obtain the amount of hemicellulose and
expressed in mg hemicellulose g-1 dry wt.
Lignin was estimated according to the procedure
of Thimmaiah (1999). For lignin estimation 100 mg
oven dried sample was moisten in a mortar with
water, grinded with ether until it was free from
chlorophyll pigment. The sample was centrifuged at
2000 g for 5 min. Supernatant was discarded and
the pellet was washed with water. The process was
repeated twice. Two ml of NaOH was added to the
residue and placed it for 12 -16 h at 70-80° C. The
hot alkali extract (0.55 ml) was taken in a test tube
containing 1-3 μg of phenol, 0.4 ml of (0.5 M) Tris-
HCl (pH 9.0) buffer and 0.05 ml freshly prepared
alcohol solution containing 25 μg of 2,6 –
dichloroquinone chlorimide. After incubation for 1 h
in the room temperature the absorbance was taken
at 610 nm using guaiacol as a standard. By using the
conversion factor 32 the lignin content was
calculate (32 x mg phenol = mg lignin)