Materials and Methods
Plant materials and preparation of aqueous, ethanol and
n-butanol fractions
Fresh healthy aerial parts of Tragia hispida ,Justicia adathoda and
tubers of Cyperus rotundus were collected from Matara District,
in the Southern Province of Sri Lanka during July to September 2017.J. adathoda and tubers of C. rotundus were authenticated
by Nawinna Ayurveda research institute. The T.hispida plant was
authenticated by the National Herbarium, Royal Botanical Garden,
Peradeniya, Sri Lanka and a voucher specimen (Th/Matara/01), was
deposited in the National Herbarium, Royal botanical garden, Peradeniya,
Sri Lanka.
Fresh aerial parts (leaves and stems) of T. hispida , dried tubers
of Cyperus rotundus and fresh aerial parts of Justicia
adathoda were washed, air dried and subsequently oven dried at 400C for 4 days and ground using a mechanical grinder.
Two 300 g samples of T. hispida were extracted separately by
refluxing the plant material for 2 hours in either 1.5 L of distilled
water and 80 % ethanol to obtain an aqueous extract (THA) and an
ethanol extract (THE) respectively. A portion of the THA extract was
freeze dried and a portion of it was mixed with n-butanol to to obtain
the butanol soluble fraction of the
aqueous extract of T. hispida (THB).
In the preparation of C. rotundus plant extract, two 50 g samples
of C. rotundus were
extracted separately by refluxing the plant material for 2 hours in
either 250 mL of distilled water and 80 % ethanol to obtain an aqueous
extract (CRA) and an ethanol extract (CRE) respectively. CRE was
evaporated under a vacuum to obtain a sticky brown semi-solid compound.
The aqueous extract (CRA) was freeze-dried for 5 days to obtain a light
brown coloured powder. Justicia adathoda, was prepared similarly
with two 50 g samples extracted separately by 250 mL each of distilled
water and 80 % ethanol by refluxing for 2 hours. Then 80 % ethanol
extract (JAE) was evaporated under a vacuum to obtain brown semi-solid
compound. The aqueous extract (JAA) was freeze-dried for 5 days to
obtain a yellowish brown coloured powder.