Cytotoxic Activity of Selected Medicinal Plants from Papua,

Indonesia

Septriyanto Dirgantara

, Rosye H. R.Tanjung

, Rahmawati Nurlatifah

and

Edy Meiyanto

Department of Pharmacy, Faculty of Mathematic and Natural Science, Cenderawasih University, Jayapura

Department

Biology, Faculty of Mathematic and Natural Science, Cenderawasih University, Jayapura ;

Cancer Chemopreventive Research Center (CCRC), Faculty of Pharmacy,

Gadjah Mada University, Yogyakarta

Keywords: Cytotoxic, BSLT, Medicinal Plants, Papua.

Abstract: Some medicinal plants from Papua which have been used traditionally as anticancer agents for many ethnic

in Papua Islands. But, scientific evidence for this study were still limited. Objective: The present study was

conducted to test for in vitro Brine Shrimp Lethality Test (BSLT) of the seven selected medicinal plants from

Papua, namely, Drymis piperita (local name : Kayu Akway), Myrmecodia beccarii (Sarang semut),

Biophytum petersianum (Rumput kebar), Vernonia amygdalina (Daun Afrika),Villebrunea rubescens (Daun

Jilat), Laportea aestuans (Daun Gatal) and Breynia cernua (Katuk hutan). Seven ethanolic extracts were

evaluated for their cytotoxic activity against Artemia salina Leach with concentration extracts from 10;100;

and 1000 ⎧g/mL. Cytotoxicity was evaluated in terms of LC

value from the 24 hour counts using the probit

analysis method. Results: Ethanolic extracts of Myrmecodia beccarii showed the highest cytotoxicity with

was 8.33 ⎧g/mL against brine shrimp, but all selected medicinal plants showed the active potential

cytotoxic activity because the LC

value were <1,000 ⎧g/mL. From this research, M.beccarii indicated the

possible potential use of medicinal plants from Papua as anticancer agents.

1 INTRODUCTION

Papua Province is rich in biodiversity. The

indigenous Papuans utilize most of endemic plants

as food, board and medicines. Several Papua

endemic plants such as Drymis piperita,

Myrmecodia beccarii, Biophytum petersianum,

Vernonia amygdalina Villebrunea rubescens,

Laportea aestuans and Breynia cernua often used as

a medicinal plant for the Papuan people (Dinkes

Papua, 2016)

Drymis piperita Hook.f. (local name : Kayu Akway)

is a woody, evergreen and aromatic plan that was a

member of Winteraceae. This plant is used by Sougb

tribe living in Sururey village, District of

Manokwari, West Papua to heal malaria and to

enhance the vitality of body (Cepeda, 2015).

Myrmecodia beccarii known as “sarang semut”

(Rubiaceae) from Merauke Region, Papua which

have been used traditionally for human vitality

enhancer, muscle pain, inflammation, and

antioxidants for supporting anticancer agents from

natural products (Dirgantara, 2013).

Biophytum petersianum (Oxalidaceae) commonly

known as “rumput kebar”, is valuable medicinal

plant from Kebar District from Manokwari Region

and the whole plant has been used in mouthwashes,

antidotes and laxatives (Sambodo, 2018).

Laportea aestuans (Urticaceae) is an indigenous

plant of Papua which has been widely used for pain

relief as traditional medication for Papua community

and the local name is “daun gatal” (Simaremare,

2017).

Villebrunea rubescens as original plants, known as

“Lick Leaves” in English and “Daun Jilat” in the

Papua local name, of the family urticaceae is a

medicinal herb with a long history of used. V.

rubescens has been used for the treatment of pain

Dirgantara, S., Tanjung, R., Nurlatifah, R. and Meiyanto, E.

Cytotoxic Activity of Selected Medicinal Plants from Papua, Indonesia.

DOI: 10.5220/0008359001550158

In Proceedings of BROMO Conference (BROMO 2018), pages 155-158

ISBN: 978-989-758-347-6

155

relieved or bruised in the local community widely.

These leaves are picked, and warmed over a fire and

then attached into the part of body pained and licked

it will give the effect of bleeding from the body. The

local people believed that the body felt better after

used the leaves (Gunawan, 2018).

Vernonia amygdalina (local name Daun Afrika)

from Asteraceae family has been used for Papua

community to medicate malaria epidemic and blood

sugar disease (Handjojo, 2018).

Breynia cernua (local name Katuk Hutan) is an

Indonesian medicinal plant (family: Euphorbiaceae)

originated from Papua which have been used

traditionally as alternative treatment for breast and

cervical cancer (Dirgantara, 2018).

Typically any research on natural substances

suspected of potentially as drugs or empirically has

been used by the community as a drug, beginning

with a pre-clinical test of toxicity to predict its safety

level, followed by other pharmacological tests.

Toxicity test method can be done in vitro and in

vivo. One of the most commonly used methods of in

vitro toxicity is the Brine Shrimp Letality Test

(BSLT) method. BSLT method is one of the fastest

and cheapest way to screen for toxicity from plant

extract by using marine animals namely A.salina

shrimp larvae. Toxicity test with BSLT method has a

broad spectrum of pharmacological activities, simple

procedure, fast and does not require a large cost, and

the results can be accounted for. In addition, this

method is often associated with a method of

screening anticancer compounds. For these reasons,

this test is best used in the initiation of natural

materials research for anticancer agents (Meyer et

al., 1982).

2 MATERIAL AND METHODS

2.1 Sample Preparation

Plant material such as Drymis piperita, Myrmecodia

beccarii, Biophytum petersianum, Vernonia

amygdalina, Villebrunea rubescens, Laportea

aestuans and Breynia cernua were collected from

some public forests in Papua and West Papua

Province, Manokwari; Merauke: Sentani; Serui and

Jayapura City. All plants were washed with

running tap water and then rinsed by distilled

water to remove any adsorbed contaminant from

sample surface.The cleaned sample was dried and

mashed to the ground to fine powder 30 mesh.

2.2 Extraction of Plant Material

Fine powder 100 gram of plant material was

macerated with 300 mL 96% ethanol at room

temperature for 3 x 24 hours, repeated until

maceration obtained translucent color. Extract of

obtained was concentrated to obtain further

combined and filtered using filter paper, and then the

solvent removed using a rotary vacuum evaporator

at 40 °C to obtain a concentrated ethanol extract of

Drymis piperita, Myrmecodia beccarii, Biophytum

petersianum, Vernonia amygdalina Villebrunea

rubescens, Laportea aestuans and Breynia cernua.

2.3 Toxicity Tests Using the Method of

Brine Shrimp Lethallity Test (BSLT)

Method of Meyer (1982), is used to study the

toxicity of the general sample using shrimp eggs (A.

salina Leach). Brine Shrimp Lethality Test (BSLT)

is one of the methods bioactive compounds present

in natural materials using shrimp larvae (A. salina).

Known toxicity properties based on the number of

larvae mortality (Mc Laughlin, 1983). An extract is

said to be toxic to A. salina if it has a value of LC50

(lethal concentration to 50% larval shrimp) less than

1,000 µg/ml.

2.4 The Hatching of Shrimp larvae

Prepared shrimp vessel for hatching eggs which

have been filled with sea water 1,500 ml, with pH of

7.0, place the lamp to warm temperatures in vessel

of hatching and fed air by using the aerator. Inserted

into the sea water of 4.0 g shrimp eggs for hatching.

Vessel hatching eggs covered with aluminum foil,

and the lights turned on for 48 hours to incubate the

eggs. After 48 hours of shrimp eggs will hatch into

larvae and ready for use. Shrimp larvae that will be

used for testing were taken using a pipette.

2.5 Preparation of Sample Solution that

will be Tested

Ethanol extract of Drymis piperita, Myrmecodia

beccarii, Biophytum petersianum, Vernonia

amygdalina Villebrunea rubescens, Laportea

aestuans and Breynia cernua that will be made

alternative dilution procedure developed by

McLaughlin et al (1983) were adopted in the

preparation of the different dilutions of the plant

extracts for BSLT where 20 mg of each extract was

dissolved in 2 mL of the solvent. The final

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

156

concentrations were 0, 10, 100, and 1000 μg/mL in

sea water. When the ethanol extract insoluble added

2 drops of DMSO (dimethyl sulfoxide 0.2%). There

were three (3) replicates in each concentration. A

control test was also prepared.

2.6 Procedure of Toxicity Test Methods

using BSLT

The seawater was put in a small plastic container

(hatching chamber) with a partition for dark

(covered) and light areas. Shrimp eggs were added

into the dark side of the chamber while the lamp

above the other side (light) will attract the hatched

shrimp. Two days were allowed for the shrimp to

hatch and mature as nauplii (larva). After two days,

when the shrimp larvae are ready, 4 mL of the

artificial seawater was added to each test tube and 10

brine shrimps were introduced into each tube. Thus,

there were a total of 30 shrimps per dilution. Then

the volume was adjusted with artificial seawater up

to 5 mL per test tube. The test tubes were left

uncovered under the lamp. The number of surviving

shrimps were counted and recorded after 24 hours.

Using probit analysis, the lethality concentration

(LC

) was assessed at 95% confidence intervals.

of less than 100 μg/mL was considered as

potent (active). As mentioned by Meyer and others,

value of less than 1,000 µg/mL is toxic while

value of greater than 1,000 µg/mL is non-toxic.

The percentage mortality (%M) was also calculated

by dividing the number of dead nauplii by the total

number, and then multiplied by 100%. This is to

ensure that the death (mortality) of the nauplii is

attributed to the bioactive compounds present in the

plant extracts.

Observations were made after 24 hours to calculate

the percentage of mortality shrimp larvae A. salina.

Mortality data are used to calculate the value of

Lethal Concentration 50 (LC

). After 24 h of

incubation, the vials were observed using a

magnifying glass and the number of survivors in

each vial was counted and noted. From this data,

the percentage of mortality of the nauplii was

calculated for each concentration and LC

values

with 95% confidence limits were determined using

Probit analysis Finney (Meyer, 1982).

3 RESULTS AND DISCUSSION

The Brine Shrimp Lethality Test (BSLT)

represents a rapid, inexpensive and simple bioassay

for testing plant extracts bioactivity which in most

cases correlates reasonably well with cytotoxicity

and anti-tumor properties (Krishnaraju, 2005).

Presently there is an increasing interest world

wide in herbal medicines accompanied by

increased laboratory investigation into the

pharmacological properties of the bioactive

ingredients and their ability to treat various

diseases (Lobo, 2009).

Toxicity tests of crude ethanol extract of Drymis

piperita, Myrmecodia beccarii, Biophytum

petersianum, Vernonia amygdalina Villebrunea

rubescens, Laportea aestuans and Breynia cernua

conducted to determine the level of toxicity of the

extracts against larvae shrimp A. Salina. The test

results showed that seven ethanolic extracts from

Papua were potent cytotoxic activity because the

value less than 1,000 µg/mL that at different

concentration levels will have an impact on

mortality and larval toxicity of this case is shown in

Table 1.

Based on results, mortality of A. salina in ethanol

extract of plant M. beccarii showed the highest

potential cytotoxic activity with low LC

value with

8.33 µg/mL with minimum concentration (10

µg/mL) can reach 50% mortality after 24 hours of

treatment. Mortality of A.salina in ethanol extract of

L. aestuans, V. rubescens and D. piperita showed

high mortality with LC

34.95; 61.82 and 79.59

µg/mL respectively. Then, the cytotoxic activity for

B.cernua (255.76 µg/mL), B. petersianum (463.61

µg/mL) and V.amygdalina (865.58 µg/mL) showed

active cytotoxic effect with LC

more higher than

another plant extracts. But, all plants extracts from

this research showed the active potential activity

Toxicity testing results of crude extracts showed the

percentage of A. salina larvae mortality increased

along with the increase in concentration of the

extract. The results reveal that crude ethanol plant

extract of M.beccarii showed that the compound

contained therein are active and possess a high

bioactivity, which means that at low concentrations

has toxic and lethal larvae of A. salina. M.beccarii

has been reported to have high potential antioxidant

activity (Dirgantara, 2013) with several active

compunds such as flavonoid, tanin and

triterpenoid/steroid (Dirgantara, 2015).

Results from this study indicate that while plant

species with LC

values < 1000 μg/ml may used for

local wisdom community, this study calls for further

work aimed at isolating the cytotoxic compounds

responsible for the observed activity and to search

new compound for anticancer therapy.

Cytotoxic Activity of Selected Medicinal Plants from Papua, Indonesia

157

4 CONCLUSION

From this research, M.beccarii showed the highest

cytotoxic activity with LC

8.33 µg/mL and

indicated the possible potential use of medicinal

plants from Papua as anticancer agents.

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