Cytotoxic Activity of Crude Extract

from Premna odorata (Blanco)

Using Brine Shrimp (Artemia salina)
Assay
Cris Arn Baltazar

Kristine Joan DA. Barredo

Research Adviser

According to the Department of Health of the Philippines, cancer is

the third leading cause of mortality and morbidity next to diseases of
the heart and diseases of the vascular system (DOH, 2009). Screening
of medicinal plants and isolation of natural products provide
pharmacologically active compounds against cancer.
Alagaw is a small hairy tree with broad, rounded, or somewhat heart-
shaped base, and pointed tips. Studies have shown antimicrobial,
cardiotonic, anticoagulant, hepatoprotective, antitubercular,
antitumor properties.
This study aimed to evaluate the cytotoxic activities of crude extract
from P. odorata leaves using brine shrimp assay. Based on the results,
the crude extract from P. odorata exhibited significant cytotoxic
activities with the LC50 values of 4.68, 4.79 and 6.61 μg/ml
respectively. This means that the crude extract of the plant
compounds is promisingly cytotoxic and have anticancer or
antitumor activity. Further studies are required to investigate these
plant samples as anticancer or antitumor agents.

Extraction of crude extracts Cytotoxicity Assay

The researcher used small cups as containers for each concen-

The terrestrial plants P. odorata was collected
tration of plant sample. The cups must be labeled and arrange ran-
from the school herbarium. The plants were air dried
domly following the layout of complete randomized design (CRD).
at room temperature after collection. The dried sam-
Put in the cups the different amount of the plant extract (0 to 10 mg).
ples were crushed by using a blender. Dried and
This could be done by dissolving 100mg of the plant sample in 5 ml
crushed samples were soaked in 100% ethanol for
filtered sea water or artificial sea water. Each drop should contain 1
three days and then filtered by using filter paper. The
mg of the plant sample. This can be done by calibrating 1 ml dropper
filtrate (crude ethanolic fraction) was concentrated
to yield 20 drops. For the 0mg, put 1 drop of sea water to the con-
and reserved for solvent partitioning.
tainer.

In this bioassay, the crude extract

from P. odorata exhibited significant

cytotoxic activities with the LC50 values of

4.68, 4.79 and 6.61 μg/ml respectively.

This means that the crude extract of the

plant compounds is promisingly cytotoxic

and have anticancer or antitumor activity.

Alagaw is a small hairy tree, 3 to 8 meters high. Leaves are ovate to broadly
ovate, 10 to 20 centimeters long, with broad, rounded, or somewhat heart-
shaped base, and pointed tips. Under surface of the blade is usually covered
with short hairs, aromatic when crushed. Flowers are greenish-white or nearly
white, 4 to 5 millimeters long and borne on terminal inflorescences (cymes) 8 to
20 centimeters in diameter. Fruit is fleshy, dark purple, rounded, about 5 millime-
ters in diameter. Studies have shown antimicrobial, cardiotonic, anticoagulant,
hepatoprotective, antitubercular, antitumor properties.
Since most active plant principles are toxic at elevated doses, a possible ap-
proach to developing an effective general bioassay might simply to screen for
substances that are toxic to zoologic systems. Desiring a rapid, inexpensive, in-
house, bioassay for screening and monitoring of physiologically active plant ex-
tracts, the researcher opted to use brine shrimp (Artemia salina) as the general
bioassay tool.

In summary, crude extract of the Premna odorata

(Blanco) is promisingly cytotoxic and have anticancer or
antitumor activity. This is in parallel to the study of Cheng
Yi Ong et al, 2009, where he screened 155 extracts from
93 species of plants in Malaysia for in vitro photo-cytotoxic
activity using a human leukemia cell line, P odorata was
one of 29 plants that was able to reduce in vitro cell
viability by more than 50% when exposed to broad
spectrum light. Study of Vadivu et al (2009) also showed
the alcoholic extract with significant hepatoprotective
activity evidenced by decrease of serum enzymes,
bilirubin and lipid peroxidation, comparable to drug
silymarin. It also exhibited significant in-vitro cytotoxic
activity. Results showed the alcoholic extract not only as
an effective hepatoprotective agent, but with also
significant antitumor activity. Further studies are required
to investigate these plant samples as anticancer or
antitumor agents.