Method
for the estimation of Phaseolin content
Reagent
Required:
Methanol L.R. Grade, Sulphuric acid, Ammonia
solution.
Glass
wares and Apparatus Required:
250mL Conical flask, Gooch crucible, 50mL
Stoppered cylinder and Evaporating dish.
Procedure:
Weigh accurately about 5.0gm of dry extract powder
in a 250mL conical flask, Add about 100mL distilled water. Shake well for four hours. Keep the
suspension over night. Next day centrifuge the liquid and take 50mL supernant
liquid. Transfer the liquid into a separator & extract successively with 50mL
each of chloroform & ethyl acetate & discard the organic layer. Filter
the aqueous layer through whatman filter paper. Filtrate is slowly added with
constant stirring to 100mL of alcohol into a preweighed dish. Left overnight to
get precipitate. Decant the supernatant liquid & discard. Dry the
precipitate at 105°C to constant weight
& weigh.
Calculation:
Percentage of Phaseolin
content is calculated as:
(A – B) X 100
=
-------------------------X 100
W X 50
A = Weight of residue and dish
B = Weight of empty dish
W = Weight of material taken
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