Journal Basic Science And Technology, 1(2),27-29,2012

ISSN : 2089-8185

Phytochemical screening and Antibacterial

activity of methanol extract of mangrove plant
(Rhyzophora mucronata) from Porong River
Estuary
Rahmi Nurdiani, Muhamad Firdaus, Asep Awaludin Prihanto*
Dept. Fishery Product Technology, Faculty of Fisheries and Marine Science, Malang, Indonesia
asep_awa@ub.ac.id

Abstract—The aims of this study were to obtain phytochemical
content and to examine antibacterial activity potency of methanol
extract of Rhyzophora mucronata. The phytochemical screening
was assayed by qualitative analysis and the antibacterial assay
determined by disk diffusion method. The results of
phytochemical analysis showed that all methanol extract of root,
bark, leaf, fruit and flower of R. mucronata contained alkaloid,
tannin, and flavonoid, in addition fruit contained steroid.,
respectively. The antibacterial activity in gram positive bacteria
(Staphylococcus aureus) and gram negative bacteria (Escherichia
coli) was showed by almost all extracts of root, bark, wood,
flower and leaf of R. mucronata.
Keywords- Phytochemical, Antibacterial, methanol extrac,
Rhyzophora mucronata.
I. INTRODUCTION
Infectious diseases are the world's leading cause of
premature deaths [1]. Furthermore, various hard work have
been made to discover new antimicrobial compounds from
many kinds of natural sources such as plants, animals, fungi,
bacteria and other microorganism. It is because the
phenomenon that many pathogenic bacteria become resistant
to antibiotics. In addition, many synthetic antibiotics smoothly
confirm the terrible effect in term of health and bacterial
resistance. Contrary to the synthetic antibiotic, antimicrobials
of plant origin are not associated with many side effects and
have an enormous therapeutic potential to heal many
infectious diseases [2].
This fact has been triggering many scientists from the
entire world to search new sources for handling it. One of
such resources is plants which are based on folk medicine.
Mangrove is well known as a coastal folk medicine for many
centuries. Mangroves have been widely used for several
human remedies. Rhyzophora mucronata is one of mangroves
that have been utilized traditionally for that purpose [3]
One of the possible methodologies that can be used for the
discovery of antibacterial is the screening of selected plant
extracts for the activity followed by bioassay. The Porong
River carried enormous materials from land to sea.
Furthermore, for more than six years it was used for drain of
waste disposal of Porong’s mud disaster due to inadequate of
oil mining by PT. Lapindo Brantas Inc. We assume that this
condition have been not only affecting the ecology but also
affecting the synthesis of the bioactive in the mangrove plants.
This study has been dedicated to investigate the
Phytochemical and antibacterial activity of extracts from
mangrove plant (R. mucronata).
II. METHODS
A. Plant collection
The mangrove R. mucronata was collected from Porong’s
river estuary. All parts (Root, bark, leaf, fruit, flower) of R.
mucronata then were packed in Poly ethylene plastic and
eventually brought to laboratory. The parts were sun dried until
± 15% water.
B. Extraction preparation
Every sample was extracted by maceration method. The 50
g dried fruit husk was soaked in 200 ml methanol in an 500
mL erlenmeyer flask for 24 hour. The extracts were then
separated from the debris by filtration with a Whatman no 1.
Afterward the extracts were concentrated using rotary
vacuum evaporator. The extracts then diluted to obtain several
concentrations of crude methanol extract. The methanol
extract were subjected for antibacterial activity.
C. Phytochemical screening
Fresh parts of R. mucronata were air dried and then
homogenized to fine powder and stored in airtight bottles at
4oC. Qualitative phytochemical analysis of the flower powder
was analyzed using Harborne method [5] and Trease and
Evans methods [6].
1. Alkaloid: 5g of each sample was weighed into a 250ml
beaker and 200ml of 10% acetic acid in ethanol was added,
covered and allowed to stand for 48 h. After filtration, the
extracts were concentrated on a water bath to ¼ of the original
volume. Concentrated ammonium hydroxide was added in
drops to the extract until the precipitation was collected,
washed with dilute ammonium hydroxide and then filtered.

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 Journal Basic Science And Technology, 1(2),27-29,2012
ISSN : 2089-8185
The residue obtained is the alkaloid, and was dried and III. RESULT AND DISCUSSION
weighed.
2. Tannins: 0.5g of each powered samples were boiled in A. Phytocemical screening
20ml of water in a test tube and then filtered. Few drops of The results of the phytochemical analysis of the R.
0.1% ferric chloride was added and observed for brownish mucronata are presented in Table 1.
green or blue black colour. The leaves contain all of the screened phytocemical. It is
3. Saponin: 25g each of the powdered samples were considered as the most abundant phytochemical compound
boiled in 25ml of distilled water in a water bath and filtered. among the others parts. However, two phytocemical is only in
10ml of the filtrate was mixed with 5ml of distilled water and
small amount namely tannin and saponin. In contrast, the root
shaken vigorously for a stable persistent froth. The frothing
presents relatively few quantities of phytochemical
was mixed with 3 drops of olive oil and shaken vigorously,
then observed for the formation of emulsion. compounds. Even though, its alkaloid was as extremely
4. Phenolic: 10 g sample was boiled with 50 ml of ether positive as the leaves.
for the extraction of the phenolic component for 15 min. 5 ml
TABLE I. PHYTOCHEMICAL CONTENT
of the extract was pipetted into a 50 ml flask, and then 10 ml
of distilled water was added. 2 ml of ammonium hydroxide Phytochemical Parts of mangrove (R. mucronata)
solution and 5 ml of concentrated amylalcohol were also Screening Leaves Root Bark Fruit Flower
added. The samples were made up to mark and left to react for Alkaloid ++++ ++++ ++++ +++ ++++
30 min for colour development. This was measured at 505 Tannin + + ++++ ++ +++
Saponin + + +++ ++ +++
nm. Phenolic ++++ - ++++ ++ ++
5. Flavonoids: A portion of the powdered plant samples Flavonoid ++++ - ++++ + ++
were separately heated with 10ml of ethyl acetate in a water Terpenoid ++++ - ++++ + +
bath for 3min. The mixtures were filtered and 4ml of each Steroid ++ + - - -
filtrate were shaken with 1ml of dilute ammonia solution. A Glycosides ++++ + +++ ++ +++
(-)=Negative; (+)=Weak positive; (++)= Positive (+++)=strong positive; =Extremely positive
yellow colour observation indicates the presence of
flavonoids.
6. Terpenoid: 5 ml of each extract was mixed in 2 ml of This data illustrate that bark of R. mucronata was second
chloroform and Concentrated H2SO4 (3ml) was carefully highest amount and most diverse phytochemical (alkaloid,
added to form a layer. A reddish brown colouration of the tannin, saponin, phenolic, flavonoid, triterpenoid and
interface was formed to show positive result for the presence glycosides ). This obtained result confirms to the findings of
of terpenoid. [8]. The bioactive compounds on the medicinal plants
7. Steroids: 2ml of acetic anhydride was added to 0.5g employed contain various secondary metabolites such as
ethanol extract of each sample with the addition of 2ml phenols, tannins, alkaloids, flavonoids, steroids and glycosides
H2SO4. A colour change from violet to blue or green indicates in appreciable quantities
the presence of steroids. All metabolites have been shown to be responsible for
8. Glycosides: 5 ml of each extract was treated with 2 ml therapeutic activity of plants [9]. Bandaranayake [3] listed the
of glacial acetic acid containing one drop of ferric chloride bioactivities of the R. mucronata (Table 2). From his result, we
solution. This was underlayed with 1 ml of. H2SO4. A brown predict that the antibacterial activity will be detected in leaves
ring of the interface indicates a deoxy sugar characteristic of and bark.
Cardenolides. A violet ring may appear below the brown ring,
while in the acetic acid layer, a greenish ring may form just TABLE II. TRADITIONAL USES, CHEMICAL CONSTITUENTS AND ACTIVITIES
gradually throughout thin layer. OF R. MUCRONATA

D. Antibacterial assay Botanical Traditional

Tested for Chemical compounds
name use/properties
The antibacterial activity test was done by paper disc
R. treatment of Antiviral, anti- alkaloids,
diffusion method [7]. The microorganisms used in this study mucronata elephantiasi,hae HIV, growth Anthocyanidins,
were Staphylococcus aureus and Eschericia coli. The 108 matoma, hormone tests carbohydrates,
cfu/ml of each inoculums were seeded in Mueller Hinton Agar hepatitis, ulcers, on plants, carotenoids, chlorophyll
and a febrifuge, biotoxicity on a, b, a & b, condensed
(MHA, Oxoid) using sterile cotton swab. Sterile 6 mm paper (B,Fl, Fr, L, R) fingerlings of and hydrolysable
blank discs were impregnated with different concentration of fish, (B, Fr,L, tannins, gibberellins,
crude extract and plated onto seeded Petri disc. The plates S, Fl, W, R) flavonoids,inositols,
lipids, minerals,
were then incubated in 350C for 24 hour. The inhibition zone polysaccharides,
around each disc was measured in millimeter polyphenols,
procyanidins, proteins,
saponins, steroid,
riterpenes, (B, L,R, S)
B=bark; L=leaves; Fr=fruits; R=roots; St=stems; Fl=flower; Sd=seed; W=whole plant; Lt=latex

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 Journal Basic Science And Technology, 1(2),27-29,2012
ISSN : 2089-8185
B. Antibacterial activity
In this study illustrated that almost all parts of the R.
mucronata inhibited the growth of all bacterial strains, but
their effectiveness varied (Table 3).
TABLE III. INHIBITION ZONE (MM) OF R. MUCRONATA
EXTRACTS.
Tested bacterium
Parts of mangrove
S. aureus E. coli
Leaves 6.1 mm 6.4 mm
Root 7.6 mm -
Bark 8.8 mm 8.6 mm
Fruit 7.1 mm 6.2 mm
Flower 7.5 mm 7.1 mm
R. mucronata bark extract gave the highest activity in both
tested bacteria with inhibition zone value of 8.8 mm and 8.6
mm against S. aureus and E. coli, respectively. R. mucronata
inhibited S. aureus dan E. coli due to the substances of each
parts. For instance, alkaloid, tannin, saponin, phenolic,
flavonoid, triterpenoid, and glycosides.
Study by Joel and boemba [10] indicated the same result
with this result. The leaves of R. mucronata that was collected
from Tamilnadu exposed antibacterial activity on both S.
aureus and E. coli. Flavonoid is well known as secondary
metabolite that has antimicrobial activity [11]. The extract of
Rhizophora mucronata bark is used for controlling diarrhea.
The anti- diarrhea of the extract is commonly due to the
bactericidal of the extract [12].
The bark extracts of R. mucronata displayed extensively a
competitive inhibitory potency with effective flower extracts
of R. mucronata on the tested isolates which majority are
Gram positive bacteria (S. aureus) known for their ability to
form resistance to drug. The mangrove parts seemed effective
on all the bacterial isolates. Conversely, the root is clearly
ineffective in E. coli. The variations in inhibitory potency
because of variations in the secondary metabolites
concentrations in the plant parts.
This finding seemingly related to mangrove habitat that
have been contaminating by Lumpur lapindo waste for six
year . In some cases, a ‘secondary’ metabolite may be
essential for survival under particular environmental
conditions Based on previous tudies that examined the mud
have found low levels of potentially toxic heavy metals and
organic contaminants [13]. In addition, Waste from the drill of
oil from east java, commonly contains Arsenic, Barium,
Cadmium, Chromium, Copper, Lead, Mercury, Selenium,
Silver, Zinc [14].
IV. CONCLUSION
Only the leaves which have diverse phytochemical
compounds, even though, the present of tannin and saponin is
weak positive. The parts of R. mucronata investigated in this
study showed some antibacterial activity against S. aureus and
E. coli. It is interesting to note that just about all parts showed
broad spectrum antibacterial activity. These promissory
extracts open the possibility of findings new clinically
effective antibacterial compounds from Indonesian natural
resources.
ACKNOWLEDGMENT
This research was partly funded by World Class University
Grant, Brawijaya University, Directorate General of Higher
Education, Ministry of Education, Indonesia
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