26101-7979 IJBAS-IJENS © February 2012 IJENS ANTIMICROBIAL AND IDENTIFICATION OF ACTIVE COMPOUND

International Journal of Basic & Applied Sciences IJBAS-IJENS Vol: 12 No: 01 69

126101-7979 IJBAS-IJENS © February 2012 IJENS I J E N S

ANTIMICROBIAL AND IDENTIFICATION OF ACTIVE COMPOUND

Curcuma xanthorrhiza Roxb.

Wibowo Mangunwardoyo

1), Deasywaty

2) , Tepy Usia

2)

1)Department of Biology, Faculty of Mathematic and Sciences, University of Indonesia,

Depok, 16424. Indonesia.

2) Center of Food and Drug Research, National Agency of Drug and Food Control,

Jalan Percetakan Negara 23, Jakarta, Indonesia.

Correspondence: [email protected] ; [email protected]

Abstract

The ethanol 70% extract of Curcuma xanthorrhiza Roxb inhibited the growth of Gram positive bacteria

S. aureus and S. mutans in a concentration of 1.0-5.0% (w/v), while B. cereus in a concentration of 2.0-5.0%

(w/v). The Minimum Inhibitory Concentration (MIC) of ethanol 70% extract toward S. aureus and S. mutans

were 0.1% (w/v), while against B. cereus it showed 2.0% (w/v). Phytochemistry analysis of ethanol 70% extract

consists of alkaloid, quinone, and terpenoids. Thin Layer Chromatography (TLC) analysis showed five spots,

the third spot effectively inhibited Staphylococcus aureus ATCC 25923, Streptococcus mutans type F (MUI)

and Bacillus cereus ATCC 11778 and contain terpenoids. Analysis UV-Vis and Infra Red spectrophotometry

showed that it contains phenolic group at absorbance 275.2 nm and have hydroxyl (-OH) and carbonyl (C-O)

functional groups. The result of GC-MS indicated that the extract which contains one compound is

xanthorrhizol m/z 218.

Index term: antimicrobial; Curcuma xanthorrhiza Roxb.; GC-MS; TLC; UV-Vis and infra-red

spectrophotometry.

I.INTRODUCTION

Curcuma xanthorrhiza Roxb is one of many medicinal plants that had been used for generations in

Indonesia [1]-[2]-[3]. Most people use the rhizome of this plant as they believe it has medicinal effect. The

rhizomes of Curcuma xanthorrhiza Roxb contain volatile oil, saponin, flavonoid and tannin [1]-[3]-[4] .

Chemistry analysis showed that the main substances of Curcuma xanthorrhiza Roxb are starch (48.18-59.64%),

fiber (2.58-4.83%), volatile oil such as, phelandren, camphor, tumerol, sineol, borneol, and xanthorrhizol (1.48-

1.63%), and also curcuminoid like, curcumine and desmetoxicurcumine (1.6-2.2%) [1]-[3] . Some research

about the chemical substances and the benefit of Curcuma xanthorrhiza Roxb had been conducted. One of those

studies found that the extract of Curcuma xanthorrhiza Roxb in ethanol 96% could inhibit the growth of

Staphylococcus aureus and Staphylococcus epidermidis as the cause of pimples [5]. Xanthorrhizol isolation

from Curcuma xanthorrhiza Roxb methanol extract could inhibit the growth of bacteria Streptococcus mutans

[6,7]. Through many studies, another benefit of Curcuma xanthorrhiza Roxb as antimicroba has also been found

[1]-[6]-[8]-[9]. The volatile oil and curcuminoid are the main substances with antimicrobial effect [1]-[2]-[6]-

[9].

Analysis method using thin layer chromatography (TLC) has been done by some researcher include.

The detachment using chloroform: methanol (9:1) in seven ethanol extract and three extract of root, stem and

leaves of Accacia (Acacia aroma Gill.) [10]. Mobile phase chloroform: methanol (99:1) was used to detach the

ethanol extract of gambier leaves (Uncaria gambir) [11]. The TLC method also has been used by [12] with

solution of n-hexan: ethyl acetate (10:1) to isolate xanthorhizol from curcuma rhizomes. Spectrophotometry

method like, UV-Vis, Infra Red dan GC-MS have been used to identified natural ingredients by some

researchers, include [13] who identified triterpenoid group in the rhizomes of Curcuma zedoaria (Berg.)

Roscoe. Reference [12] identified xanthorrhizol in it.

The purpose of research is to find out the antimicrobial effect of Curcuma xanthorrhiza Roxb. and to

identify the compound class in the extract of ethanol 70% curcuma rhizomes. The findings could support the

development of curcuma as antimicrobial element to enrich the traditional medicine in Indonesia.

mailto:[email protected]



II. MATERIAL AND METHOD

A.Plant materials

The sample for this research is dried simplicias of Curcuma xanthorrhiza Roxb rhizomes from P.T.

Vitaher, Semarang. The plants were cultivated in a high altitute about 75-100 m and were harvested after 10

months. The simplicias were dried by oven with the temperature started from 50-55º C for 7 hours.

B.Microorganisms Staphylococus aureus ATCC 25923, Bacillus cereus ATCC 11778, anaerob Streptococcus mutans

type F (MUI), Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, anaerob Porphyromonas

gingivalis ATCC 33277 and Candida albicans ATCC 10231. These microbes were supplied by Microbiology

Laboratory, Control Laboratory of Drug and Food, NA-DFC (Badan POM) and Microbiology Laboratory,

Faculty of Dentistry, University of Indonesia.

C.Media Brucella Broth was used as a medium for Porphyromonas gingivalis while TSB was used for growing

Staphylococcus aureus, Bacillus cereus, Pseudomonas aeruginosa, also Escherichia coli, and BHIB medium +

yeast extract were used for growing Streptococcus mutans.

D. Extractions An amount of 100 g powder of Curcuma xanthorrhiza Roxb was added by aquadest till it reached 500 ml then

boiled for 20 minutes [14] and was filtered by glass wool. Thick extract of aquadest was acquired by

evaporating in a waterbath. The filtered residue was reflucted by ethanol 70% within one hour and then was

filtered by glass wool. The ethanol was eliminated by evaporating with vacuum evaporator at 40º C. The

residue of ethanol was being extracted back with dichloromethane maceratively with shaker at 120 rpm for 24

hours. The extract of dichloromethane was evaporated by vacuum evaporator at 40º C until thick extract of

dichloromethane was acquired [13]-[15].

E.Antimicrobial and antifungal assay broth dilution methods The extract was dissolved by aquadest in various concentrations of 50%, 40%, 30%, 20% and 10%

(b/v). Every single tube was filled by the mixture of medium and extract in various concentrations. Then, the

bacterias were inoculated into the appropriate medium for about 200 µl with Mc Farland turbidity of 0.5 and

then were incubated at 35-37º C for 18-24 hours. The anaerob bacterias were incubated using anaerobic jar [16]-

[17].

F. Minimum Inhibitory Concentration Identification (MIC) This study used broth dilution method. The extract was dissolved until it reached concentration of

0.025%; 0.05%; 0.10%; 0.25%; 0.50% and 0.75% (b/v) The test was repeated for 3 times [16]-[17].

G.Identification Chemical Composition Using Thin Layer Chromatography (TLC) [18] Ethanol 70% extract was identified using TLC, silica gel 60F254 (20x20 cm) with 0.25 mm thickness. As the

developer solvent was n-hexan: ethyl acetate (14:1). The eluent distance was 15 cm from the spotted starting

point. The developer solvent was put into chromatography vessel, and let it till it became saturated. Then, an

amount of 10 µl extract solution was used to spot silica gel plate. Stain observation has been done under UV

light 254 nm to see fluorecensy and reactant CeSO4 spraying. Every spoted was scraped for being tested for its

activity against microbes.

H. TLC Results identification using spectrophotometry UV-Vis The TLC scraped with Rf value showing antimicrobial activity then were put into chromatography column in

the form of pipette, eluted with ethanol 70% and accumulated in vials. The results were measured by

spectrophotometer UV-Vis [19]-[20].

I. TLC Results identification using spectrophotometry Infra Red An amount of 1 mg chromatographed scraped that has been eluted with ethanol 70% and heated by waterbath

was made into a thin disc with 100 mg Potasium Bromide addition. This thin disc was observed by Infra Red

spectrophotometer [21]-[22].

J.TLC Results Identification using spectroscopy GC-MS The TLC scraped with Rf value showing antimicrobial activity then were put into chromatography

column in the form of pipette, eluted with ethanol 70% and accumulated in vials. The results were identified

using spectroscopy GC-MS to find out the compound class and its molecular weight [12]-[13]-[23].



III. RESULTS AND DISCUSIONS

A.Rhizome extraction (Curcuma xanthorrhiza Roxb.)

The result of extraction 100 g powder of Curcuma xanthorrhiza Roxb with aquadest, ethanol 70% dan

dichloromethane can be seen in the Table I.

Table I.

Yield from extraction of 100 g powder of Curcuma xanthorrhiza Roxb.

Solvent Weight of yield (g) Characteristic of yield

Aquadest 63.54 g

Yellow brownish, condensed

Ethanol 70% 13.33 g

Brown, condensed

Dichloromethane 3.01 g

Yellow brownish, condensed

The yield from aquadest has the highest result. This result could be because there are starch and phenol

compounds in the extraction and also because aquadest is a polar solvent that could dissolve starch and phenol

[18]. References [24] and [25] reported that the extraction in aquadest has second metabolite substances which

are, alkaloid, saponin and quinone. Starch is the main component of Curcuma xanthorrhiza Roxb about 48.18-

59.64% in composition [1]-[2]. The high percentage of starch is influenced by the altitude where it cultivated.

When it planted below 240 m, it will contain high percentage of starch[12].

Extraction with ethanol 70% was conducted since ethanol 70% is a versatile solvent for preliminary

extraction and it has universal charactheristic [18] so it can attract polar compounds in the rhizomes especially

alkaloid [4]-[26] curcuminoid and terpenoid [18] and also non polar compounds [27]. Another component as a

result of the extraction using ethanol is phenol compound [25]-[26]-[28]-[29].

Extraction of the rhizomes with dichloromethane has the least amount of yield, since dichloromethane

is a semi polar solvent. As a result, the extraction could only attract some of semi polar and non polar

compounds such as, some of flavonoids [30], triterpenoid [22]-[25] alkaloid [30] and saponin[25].

B.Antimicrobial activity rhizome (Curcuma xanthorrhiza Roxb.)

Rhizome extractions could inhibit the growth of all Gram positive tested as shown in the Table II.

Table II.

Antimicrobal activities in the extract of Curcuma xanthorrhiza Roxb in aquadest, ethanol 70%, and

dichloromethane against Gram positive bacteria.

Microbes

Concentration

% (b/v)

Inhibitory capacity

Aquadest

extract

Ethanol 70%

extract

Dichlorome-

thane extract

Staphylococcus

aureus

ATCC 25923

5.0 + + +

4.0 + + +

3.0 + + +

2.0 + + +

1.0 + + +

Bacillus cereus

ATCC11778

5.0 - + -

4.0 - + -

3.0 - + -

2.0 - + -

1.0 - - -

Streptococcus mutans

Type F (MUI)

5.0 + + +

4.0 + + +

3.0 + + +

2.0 + + +

1.0 + + +

Note: +: the extract could inhibit the growth of bacteria

- : the extract could not inhibit the growth of bacteria



The inhibitory process against Gram positive bacteria is possibly caused by the phenol compound in

the extraction that inhibites the growth of the microbes. High phenol penetration into the cell could cause

protein coagulation and cell membrane lysis [28]. The phenol inhibitory mechanism is by hydrogen binding

between hydroxyl group in the phenol compound and cell membrane protein, which cause membrane

permeability imbalance. This mechanism makes the essensial components of the cell leak out of the cell and

cause cell death [28]-[29]-[31] . Simple cell wall construction with no outer layer makes antibacterial compound

to permeate the cell wall and cause cell wall biosynthesis disturbance [5]- [25]- [32]-[33].

The extraction in aquadest, ethanol 70%, and dichloromethane could not inhibit the growth of Gram

negative bacteria like, E. coli, P. aeruginosa, P. gingivalis and fungi C. albicans (Table III). This could

possibly happen because the extract concentration could not penetrate the cell wall of Gram negative bacterias

and fungi. The cell wall of Gram negative bacterias consist of more complex composition than the one in Gram

positive bacterias. Not only peptidoglycan, but Gram negative bacterias also have another outer layer that

consists of lipopolysacharide, lypoprotein, and periplasm which is binded to peptidoglycan [25]-[34]

Lipopolysacharide in the outer layer has a function as a defence system with peptidoglycan and select foreign

substances. Lipoprotein contains hydrophilic protein called porin. There is a big possibility that because of

porin in the outer membrane of Gram negative bacterias, the extraction could not permeate into the cell since the

extraction is hydrophobic [25]-[34].

Table III.

Antimicrobal activity in the extract of Curcuma xanthorrhiza Roxb in aquadest, ethanol 70%, and

dichloromethaneagainst Gram negative bacteria and fungi.

Microbes

Concentration

% (b/v)

Inhibitory capacity

Aquadest

extract

Ethanol 70%

extract

Dichlorome

thane extract

Escherichia coli

ATCC 25922

5.0 - - -

4.0 - - -

3.0 - - -

2.0 - - -

1.0 - - -

Pseudomonas

aeruginosa

ATCC 27853

5.0 - - -

4.0 - - -

3.0 - - -

2.0 - - -

1.0 - - -

Porphyromonas

gingivalis

ATCC 33277

5.0 - - -

4.0 - - -

3.0 - - -

2.0 - - -

1.0 - - -

Candida albicans

ATCC 10231

5.0 - - -

4.0 - - -

3.0 - - -

2.0 - - -

1.0 - - -

Note:

(-): the extract could not inhibit the growth of bacteria

The cell wall of Gram negative bacterias has a high concentration of lipid that makes these bacterias

more resistant against chemical compound and impermeable with limited diffusion [35]. Gram negative

bacterias have more complex cell wall composition and it has non polar characteristic so all those three extracts

which contain polar to semi polar compound have more difficulty in penetrating the bacterias cell wall [28].

This also has been reported by references [15]-[36] and reference [25] mentioned that Gram negative bacterias

have resistency capacity against antimicrobes more than Gram positive bacteria.

All those three extracts were tested against C. albicans showed that all of them could not inhibit the

growth of C. albicans. This is possibly because there is not any inhibitory process since chemical substance

within the extracts could not penetrate the sterol layer in the cell wall to cease the stiffness of chitine synthesis

on cell wall [37]-[38] . Reference [39] has quoted that the fungi cell wall has a stiff characteristic and consists

of chitine, glucan dan mannan, and mainly contain 80-90% polysaccharide. Reference [36] reported that 90%



ethanol extract of Sida acuta Burm. also could not inhibit the growth of C. albicans. Also [34] reported that

berry extract in acetone water solvent (70:30) also could not inhibit C. albicans growth.

C. Minimum Inhibitory Concentration (MIC)

Minimum inhibitory concentration has been done with the extract concentration of 0.025%; 0.05%;

0.10%; 0.25%; 0.50%; and 0.75% (b/v) (Table IV).

Table IV.

The result of Minimum Inhibitory Concentration (MIC) of ethanol 70% extractagainst Staphylococcus aureus

ATCC25923 and Streptococcus mutans Type F (MUI)

Concentration

(%)

Minimum Inhibitory Concentration (MIC)

Staphylococcus aureus

ATCC 25923

Streptococcus mutans

Type F (MUI)

0.025 - -

0.05 - -

0.10 + +

0.25 + +

0.50 + +

0.75 + +

Note:

+ : Curcuma extract could inhibit the growth of tested microbes

- : Curcuma extract could not inhibit the growth of tested microbes

Ethanol 70% extract could inhibit the growth of S. aureus dan S. mutans in 0.10-0.75% (b/v)

concentration. Antimicrobial activity was affected by the concentration of extract. The higher the concentration,

the higher the inhibition capability will be. Reference [5] has reported that 95% Curcuma ethanol extract is

0.4% to S. aureus. Research of [23] on Cinnamomum iners leaves in methanol extract showed that it contains

xanthorrhizol with MIC about 0.78 mg/ml to S. aureus. Also, reference [7] has reported that the isolated

xanhorrhizol from Curcuma methanol extract (Curcuma xanthorrhiza Roxb.) could inhibit S. mutans in 5.0

µMol/l concentration.

D. Phytochemical identification of rhizome.

Classes of compound identification in ethanol 70% extract (Table V)

Table V.

Phytochemical analysis result of 70% ethanol extract (Curcuma xanthorrhiza Roxb.)

No. Classes of compound Result Characteristic

1 Alkaloid + Bouchardat: brown sediment,

Mayer: -Dragendorf: brick red sediment

2 Flavonoid -

3 Saponine -

4 Tanine -

5 Quinone + NaOH: red

6 Terpenoid/Steroid + Lieberman-Bouchardat: purple-violet

7 Glikoside -

Class of compound identification resulting negative on flavonoid identification because of the absent of

red colour, saponine was not foaming around 1-10 cm after 10 minute, tannine was not turned into blue, and

glycoside was not changed from red to blue or purple after the addition of Keller Killiani reagent [18]-[21]-[39].

E. Thin Layer Chromatography analysis.

The result of TLC ethanol 70% extract of Curcuma on 60F254 plate with n-hexan: ethyl acetate (14:1)

developer, were 5 spots with variable Rf value (Table VI).



Table VI.

The result of TLC ethanol 70% extract of curcuma with n-hexan : ethyl acetate (14:1) developer.

No.

Silica gel 60F254

Rf

Colour

254 nm CeSO4

0 Yellowish brown Yellowish brown

1 0.16 Light purple Brown


3 0.64 Dark purple Brown-purple

4 0.73 Dark purple Brown


According reference [40] Curcuma longa L. methanol extract with mobile phase chloroform: ethanol: acid

acetate (48:2:0,1) has Rf value of 0.4; 0.23 and 0.19. Ethanol 70% extract of curcuma is assumed to have some

terpenoid compounds, such as xanhorrhizol. This is supported by the existence of purple after TLC plate

sprayed by H2SO4 reagent. According to reference [18] the terpenoid compounds on TLC plate that was sprayed

with reagent will form purple colour. Reference [12] was succeed in get Rf value from curcuma rhizome

extraction with ethanol 96% solvent and developer solvent n-hexan: ethyl acetate (10:1) on Rf 0.56 and 0.86.

Reference[41] said that xanthorrhizol Rf value is 0.58. Xanthorrhizol is the main antimicrobe chemical

compound in curcuma [1]-[6]-[7]-[23]-[42].

F.Antimicrobial activity and phytochemical analysis results of TLC

Antimicrobial activity test from TLC ethanol 70% extract curcuma by dilution tube method on Gram

positive Staphylococcus aureus, Streptococcus mutans, and Bacillus cereus inhibit on the third spot with 0.64 Rf

value. The third spot on CeSO4 reagant showed terpenoid compound assumed by the purple colour. The purple

thin layer of chromatography with reagent is terpenoid [18]. Reference [12] has shown that xanthorrhizol TLC

from curcuma methanol extract with n-hexan: ethyl acetate gave spot on Rf 0.54 and 0.86. The absence of

microbe inhibition on fraction 1, 2, 4, and 5 were assumed because the fraction concentration of

chromatography is too small so that the compound in the extract could not inhibit the bacteria growth. TLC

scrape confirmation by alkaloid, quinone, and terpenoid reagent showed that Rf 0.64 contain terpenoid

compound after being tested by Lieberman and Bouchard reagent. It gave a positive result with purple violet

formation. A test using alkaloid and quinone reagent showed negative in result [18]-[21]-[39].

G. Qualitative Identification using UV-Vis, Infra Red dan GC-MS Spectrophotometry

Analysis result of active compound identification using spectrophotometry UV-Vis (Fig 1).

Fig 1. UV-Vis Spectrophotometry chromatogram profile

Identification using UV-Vis Spectrophotometry showed that the peak absorbent wave is 275.20 nm it

means that there is the electron transition from phenol compounds. Reference [43] mention that the maximum

of phenol group absorbent is in 210 – 280 nm, with electron transition of aromatic group π → π*. Reference

275.2 nm



[23] said that result that maximum absorption of Cinnamonum iners extract with Chloroform solvent is 276.0

nm, by phenol group indication. The maximum absorption ethanol extract of rhizome (Curcuma zadoria (Berg.)

Roscoe) is 242 nm [13].

Active compound identification of ethanol 70% curcuma extract by infra red Spectrophotometry test is

shown on Fig 2 and Table VII.

Fig 2. Infra red spectrophotometry profile

Table VII.

Infra Red wave numbers of ethanol 70% curcuma extract (Curcuma xanthorrhiza Roxb)

Bonds

Wave number (cm-1

)

Silverstein et al. (1963) Ethanol 70% extract

Rf 0.64

C-C, C-O, C-N 1300 - 800 1100.41

C=C, C=O, C=N, N=O 1900 - 1500 --

C=C, C=N 2300 - 2000 --

C-H, O-H, N-H 3800 - 2700 3387.06

Based on Fig 2 and Table VII we can conclude that fraction with Rf value 0.64 contains compound

with hydroxyl functional group (-OH) with wave number 3387.06 cm -1

, supported by strong absorption on

wave number 1100.41 cm-1

from C-O alcohol. Hydroxyl group (-OH) is on wave number 3400-2700 cm-1

[13]-

[43]. Reference [44] said that phenol group which has –OH dan C-O group strong absorption, has absorption

level between 3550-3200 cm-1

and C-O on wave number 1260 – 1000 cm-1

. Reference [23] identified

xanthorrhizol in Cinnamomum iners leaves extract with wave number 3382.8 cm-1

for –OH group.Reference

[12] also identified xanthorrhizol in curcuma with wave number 3400 cm-1

for hydroxyl group.

GC-MS test on 70% curcuma rhizome ethanol extract with Rf value 0.64 resulted in peak mass spectrum as

shown on Fig 3.

C-O

O-H



Fig 3. Mass spectrum of curcuma 70% ethanol extract (Curcuma xanthorrhiza Roxb.)

Based on the spectrum data, the compound on ethanol 70% extract is stated to have molecule weight 218

g/mol, and on library GC-MS data is shown to have a 99% identical characteristic with compound peak of the

ethanol 70% extract which is xanthorrhizol with 218 g/mol. Reference [41] - [45] reported that the molecular

weight of xanthorrhizol is 218 g/mol.

IV. CONCLUSION

The extraction result of 100 g rhizome powder of Curcuma xanthoriza Roxb. with ethanol 70%

aquadest solvent and dichloromethane resulting in yield with weight of 63.54 g, 13.33 g, 3.01 g. Curcuma

rhizome extract effective in inhibiting Staphylococcus aureus ATCC 25923 and Streptococcus mutans type F

(MUI) positive Gram bacteria growth, while Bacillus cereus can only be inhibited by ethanol 70%. Gram

negative bacteria consist of Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853,

Porphyromonas gingivalis ATCC33277 and Candida albicans ATCC 10231 fungi could not be inhibited by

those three extractions. Ethanol 70% extract of curcuma rhizome effectively inhibited S. aureus and S. mutans

on 1.0 – 5.0% (b/v) concentration, while B. cereus on 2.0 – 5.0% (b/v), with 0.1% (b/v) Minimum Inhibitory

Concentration (MIC) to S. aureus and S. mutan. UV-Vis and Infra Red spectrophotometry analysis showed

contains phenolic group at absorbance 275.2 nm and have hydroxyl (-OH) and carbonyl (C-O) functional

groups. Result of GC-MS indicated that the extract contains one compound which is xanthorrhizol m/z 218.

Further research on determination of molecular structure and purification of the compound should be carried

out.

REFERENCES

[1] E. Afifah. “Khasiat dan manfaat temulawak, rimpang penyembuh aneka penyakit”. Agromedika, Pustaka.

pp. 1-19. 2005.

[2] BPOM (Badan Pengawas Obat dan Makanan). “Gerakan Nasional Minum Temulawak”. Info POM vol 6,

no 6, pp 1-4. 2005.

[3] M.H. Siagian. “Temulawak sebagai tanaman obat dan budidayanya secara intensif”. Balitbang Botani,

Puslitbang Biologi LIPI, Bogor. pp. 1-8 2006.



[4] J. Tarigan, C.F. Zuhra and H. Sihotang. Skrining fitokimia tumbuhan yang digunakan oleh pedagang jamu

gendong untuk merawat kulit wajah di Kecamatan Medan Baru. J. Biologi Sumatra vol 1, no 3, pp 1-6.

2008.

[5] B. Soebagio,S.Soeryati and K. Fauziah. “Pembuatan sediaan krim antikanker ekstrak rimpang temulawak

(Curcuma xanthorhiza Roxb.)”. Prosiding Pertemuan Ilmiah Pembuatan Sediaan Temulawak

(Curcuma xanthorrhiza Roxb) dari Produk Empiris Sampai Produk Fitofarmaka, Unpad, Bandung.

2006

[6] Y. Rukayadi. Effect of xanthorrhisol on Streptococcus mutans biofilm in vitro. JurnalMikrobiologi

Indonesia vol 11, no 1, 4 p. 2006

[7] Y. Rukayadi and J.K. Hwang. In vitro activit y of xanthorrhizol against Streptococcus mutans biofilms. J.

Applied Microbiology vol 42, pp 400-404. 2006.

[8] S. Samsundari. Pengujian ekstrak temulawak dan kunyit terhadap resistensi bakteri Aeromonas

hydrophilla yang menyerang ikan mas (Cyprinus carpio). GAMMA vol 2 no1, pp 71 – 83. 2006.

[9] N. BermawieM. Rahardjo, D. Wahyuno and Ma’mun. Status teknologi budidaya dan pasca panen tanaman

kunyit dan temulawak sebagai penghasil kurkumin. Laporan Hasil Penelitian Tanaman Rempah dan

Obat. Balai Penelitian Tanaman Obat dan Aromatik, Bogor. pp. 84-97. 2008

[10] M.E Arias , J.D. Gomez, N.M. Cudmani, M.A. Vattuone and M.I. Isla. Antibacterial activity of ethanolic

and aqueous extract of Acacia aroma Gill. Life Sciences vol 75, pp. 191-202. 2004.

[11] I. Kresnawat,. and A. Zainuddin. . Aktivitas antioksidan dan antibakteri dari derivat metil ekstrak etanol

daun gambir (Uncaria gambir). Jurnal Littri vol 15, no4, p. 145.2009.

[12] D. Asriani, Isolasi xanthorrhisol dari temulawak terpilih berdasarkan nomor harapan. Tesis. Institut

Pertanian Bogor. p. 151. 2010.

[13] W.S. Rita. Isolasi, identifikasi, dan uji aktivitas antibakteri senyawa golongan triterpenoid pada rimpang

temu putih (Curcuma zedoaria (Berg.) Roscoe. Jurnal Kimia vol 4, no1, pp 20-26. 2010.

[14. S.S.H. Wijono, Isolasi dan identifikasi flavonoid pada daun katu (Sauropus androgynus (L.) Merr).

Jurnal Makara Sains vol 7, no 2, pp. 51-64. 2003.

[15] Kusmiyati and N. W. S. Agustini. Uji aktivitas senyawa antibakteri dari mikrooalga Phorphyridium

cruentum. Biodiversitas vol 8, no1, pp. 48-53. 2006.

[16] H.W. Zaenab,. Mardiastuti, V.P. Anny and B. Logawa. Uji antibakteri Siwak

(Salvadora persica Linn.) terhadap Streptococcus mutans (ATCC 31987)

dan Bacteriodes melaninogenicus. Jurnal Makara Kesehatan vol 8, no 2, pp 37-

41.2004.

[17] M.K.Oladunmuye. Comparative evalution of antimicrobial activities and phytochemical screening of two

varieties of Alcalipha wilkesiana. Intl. J. Trop. Med. vol 1, no 3, pp 134-136. 2006.

[18] J.B Harborne,. “ Phytochemical methods. A guide to modern techniques of plants analysis”. 2nd

Ed.

Chapman & Hall, London. pp.89-115. 1996.

[19] I, Batubara, T. Mitsunaga and H. Ohashi. Screening anti acne potency of Indonesian medicinal plants;

antibacterial, lipase inhibition, and antioxidant activities. J. Wood Sci. vol 55, pp. 230-235. 2009.

[20] E. K. Hayati, A.G. Fasyah and L. Saa’dah. Fraksinasi dan identifikasi senyawa tanin pada daun belimbing

(Averhoa belimbii L.). J.Kimia vol 4, no 2, pp. 193-200. 2010.

[21] E. Cahyaningsih, Identifikasi senyawa antimikroba dari herba meniran (Phyllanthus niruri L.). Tesis.

Program Studi Pasca Sarjana FMIPA. Universitas Indonesia, Depok.. 2008.

[22] I.M. Sukadana, S.R. Santi and N.K. Juliarti. Aktivitas antibakteri golongan senyawa triterpenoid dari biji

papaya (Carica papaya L.). J. Kimia vol 2, no1, pp. 15-18, 2008.

[23] F.Mustaffa, J. Indurkar, S. Ismail, M. Shah and S.M. Mansor. An antimicrobial compound isolated from

Cinnamomum iners leaves with activity against Methicillin-Resistant Staphylococcus aureus. Moleculs

vol 16, pp. 3037-3047, 2011.

[24] I. Indrawati, Potensi ekstrak air, etanol dan minyak atsiri bawang merah (Allium cepa L.) kultivar Batu

terhadap bakteri penyebab karies gigi. J. Biotika vol7, no1, pp. 40-48, 2009.

[25] S. Hidayathulla, C.K. Keshava and K.R. Chandrashekar. Phytochemical evaluation and antibacterial

activity of Pterospermum diversifolium Blume. Int. J. of Pharmacy and Pharmaceutical Sciences vol

3, no2, pp. 165-167. 2011.

[26] P.F. Omojasola, and S. Awe. The antibacterial of the leaf extract of Anacardium occidentale and

Gossypium hirsutum against some selected microorganism. Bioscience Research Communication vol

16, no1, pp. 25-28. 2004.

[27] A.Saifudin, H.V. Rahayu and H.Y. Teruna. “Standarisasi bahan obat alam”. Graha Ilmu. Yogyakarta. pp

16. 2011.

[28] T. Hertiani, S.I. Palupi, Sanliferianti and D.H. Nurwindasari. . In vitro test on antimicrobial potency

against Staphylococcus aureus, Escherichia coli, Shigella dysentriaea and Candida albicans of some

herbs traditionally used cure infection diseases. Pharmacon vol 4, no 2, pp. 89-95. 2003.



[29] K.Elfahmi, Roslan, R. Bos, O. Kayser, H.J. Woerdenbag and W.J. Quax. “Jamu. The Indonesian

Traditional Herbal Medicines”. Penerbit Eisei, Jakarta: pp. 14 – 34. 2008.

[30] Y.Fitrial, M. Astawan, S. S. Soekarto, K. G. Wiryawan, T. Wresdiyati and R. Khairina. Aktivitas

antibakteri ekstrak biji teratai (Nympaea pubescens Wild.) terhadap bakteri pathogen penyebab diare.

J. Teknol. dan Industri Pangan vol 19, no 2, pp 158-164. 2008.

[31] K.K. Al-Rubiay, N.N. Jaber, B.H. Al Mhaawe and L.K. Alrubaay. Antimicrobial of henna extract.

Oman Medical Journal vol 23, no 4, 4 p. 2008.

[32] R.J.W. Lambert, P.N. Skandamis, P.J. Coote and G.J.E. Nychas. A study of the minimum inhibitory

concentration and mode of action of oregano essential oil, thymol and carvacrol. J. Appl. Microbiol.

vol 91, no 3, pp 453-462. 20,01.

[33] A.Ajizah, Thihana and Mirhanuddin. Potensi ekstrak kayu ulin (Eusideroxylon zwageri) menghambat

pertumbuhan Staphylococcus aureus secara in vitro. Bioscientiae vol 4, no 1 pp. 37-42. 2007.

[34] L.J. Nohynek, A. Alakomi, M.P. Kähkönen, M. Heinonen, Ilkka M. Helander, Kirsi-Marja Oksman-

Caldentey, and Riitta H. Puupponen-Pimiä. Berry Phenolics: Antimicrobial Properties and Mechanisms

of Action Against Severe Human Pathogens. Nutrient and Cancer vol 54 no1, pp. 18-32. 2006.

[35] T. Hanouda, and J.R. Baker. Antimicrobial mechanism of action of surfactant lipid preparation in enteric

Gram negative bacilli. J. of. Appl. Microbiology vol 89, pp.397-403. 2000.

[36] I.E. Oboh, J.O. Akerele and O. Obasuyi. Antimicrobial activity of ethanol extract of the aerial parts of

Sida acuta Burmn. (Malvaceae). Tropical Journal Pharmaceutical Research vol 6, no 4, pp. 809-813.

2007.

[37] E. Jawetz, J.L. Melnick and E.A. Adelberg. “Medical Microbiology”. 14th

Ed. Lange Medical Publications,

Canada. pp. 6-21. 1996.

[38] T.D Brock,., M.T. Madigan, J.M. Martinko and J. Parker. “Biology of microorganisms”. Prentice- Hall

International, USA: pp. 58-66. 1994

[39] F.C. Akharaiyi. and B. Bolatito. Antibacterial and phytochemical evaluation of three medicinal plants.

J.of Natural Product vol 3, no 27-34. 2010.

[40] S. KhanI., K. Makhija,D. Khamar and S. Rani.. Development and standarization of tumeric cream by

HPLC. Int.J. Biomedical and Advance Research vol 1, no 4, pp 109-116. 2010.

[41] J.K . Hwang, J.S. Shim and Y.R. Pyun. Antibacterial activity of xanthorrhizol from curcuma xanthorrhiza

against oral pathogens. Fitoterapia vol 71, no 3, pp. 321-323. 2000.

[42] C.A.C. Arraujo. and L.L. Leon. Biological activities of Curcuma longa L. Mem Inst Oswaldo Cruz vol 96,

no 5, pp. 723-728. 2001.

[43] R.M. Silverstein, G.C. Bassler and T.C. Morril. “ Spectrometric identification of organic compounds”. 4th

Ed. John Willey & Sons, New York. pp. 104-112. 1963.

[44] J.H. Naama. A.A. Temimi and A.A. Husain. Study the anticancer activities of ethanolic curcumin extract.

African J. of Pure and Appl. Chemistry vol 4, no 5, pp 68-73. 2010.

[45] Y.H. Cheah, F.J. Nordin, R. Sarip, T.T. Tee, H.L.P. Hazihmatol, H.M. Sirat, B.A. Abd. Rasid, N.R.

Abdollah and Z. Ismail. Combined xanthorrhizol-curcumin exhibits synergistic growth inhibitory

activity via apoptosis induction in human breast cancer cells MDA-MB-231. Cancer Cell

International vol 9 no 1, pp 1-12. 2009.

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