Antibacterial activity of diphenyltin(IV) and triphenyltin(IV) 3-chlorobenzoate againts Pseudomonas aeruginosa and Bacillus subtilis

Article abstract of DOI:10.13005/ojc/330310

In this paper, we reported the syntheses and antibacterial activity test of 2 organotin(IV) compounds, diphenyltin (IV) di-3-chlorobenzoate (2) and triphenyltin (IV) 3-chlorobenzoate (4). These two compounds were prepared by the reaction of diphenyltin (IV) dihydroxide and triphenyltin (IV) hydroxide with 3-chlorobenzoic acid. These compounds were characterized by ¹H and ¹³C NMR, IR, UV-Vis spectroscopies and also based on the microanalytical data. The results of antibacterial activity by diffusion method against Pseudomonas aeruginosa and Bacillus subtilis showed that the triphenyltin(IV) 3-chlorobenzoate was active at concentration of 3.956 x 10^-4M (200 ppm), while the chloramphenicol gave inhibition of 6.1894 x 10^-4 M (200 ppm), although the halozone was bigger. This result indicated that compound 4 is potentially to be used as antibacterial substance, although the search of other derivative of organotin (IV) with other ligands is still needed to get much higher and much better activity.

Full text of DOI:10.13005/ojc/330310

ISSN: 0970-020 X
CODEN: OJCHEG
017, Vol. 33, No. (3):
Pg.1133-1139
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Free Access, Peer Reviewed Research Journal
2
www.orientjchem.org
Antibacterial Activity of Diphenyltin(IV) and Triphenyltin(IV)
3-Chlorobenzoate Againts Pseudomonas aeruginosa and
Bacillus subtilis
1
,2
1
1,
ANNISSA ,TATI SUHARTATI,YANDRI and SUTOPO HADI *
1
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Lampung,
Bandar Lampung, 35145, Indonesia.
2
Sekolah Tinggi Ilmu Kesehatan Faletehan, Serang-Banten,4216, Indonesia.
*
Corresponding author E-mail : sutopo.hadi@fmipa.unila.ac.id
http://dx.doi.org/10.13005/ojc/330310
(
Received: May 09, 2017; Accepted: May 31, 2017)
ABSTRACT
In this paper, we reported the syntheses and antibacterial activity test of 2 organotin(IV)
compounds, diphenyltin (IV) di-3-chlorobenzoate (2) and triphenyltin (IV) 3-chlorobenzoate (4).These
two compounds were prepared by the reaction of diphenyltin (IV) dihydroxide and triphenyltin (IV)
1
13
hydroxide with 3-chlorobenzoic acid. These compounds were characterized by H and C NMR,
IR, UV-Vis spectroscopies and also based on the microanalytical data. The results of antibacterial
activity by diffusion method against Pseudomonas aeruginosa and Bacillus subtilis showed that the
-4
triphenyltin(IV) 3-chlorobenzoate was active at concentration of 3.956 x 10 M (200 ppm), while the
-4
chloramphenicol gave inhibition of 6.1894 x 10 M (200 ppm), although the halozone was bigger.
This result indicated that compound 4 is potentially to be used as antibacterial substance, although
the search of other derivative of organotin (IV) with other ligands is still needed to get much higher
and much better activity.
Keywords: antibacteria, B. subtilis, organotin(IV) 3-chlorobenzoate, P. aeruginosa
INTRODUCTION
pneumonia with necrosis, light external otitis on
swimmers and eyes. B. subtilis can cause damage
on canned food which can cause gastroenteritis to
Infectious disease is one of the main
2
problems in the world health, as almost every
the consumers .
1
country faces this problem . The main cause of
infectious diseases is due to bacteria, of them are
One of the ways that can be carried out
negative gram bacteria P. aeruginosa and positive
gram bacteria B.subtilis .P.aeruginosa causes many
to inhibit the growth of bacteria in every organism
is by applying antibacterial substances .Based on
1
3
diseases such as infections of wounds, burns, bluish
green pus, urinary channel, respiratory causing the
the use of antibacterial substances, they can be
categorized as disinfectant, antiseptic, sterilizer,

1134
ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
4
-4
sanitizer . The inhibition of bacterial growth by 1.0x10 M. A Bruker AV 600 MHz NMR (600 MHz
1
13
1
antibacterial substances can be done by cell for H and 150 MHz for C) were used to obtain H
disruption mechanism which is by inhibiting its and C NMR spectra. All experiments were run in
formation or changing it after being formed, alteration DMSO-D at 298K.The number of scans used for H
13
1
6
of cytoplasm membran permeability that cause experiments were 32 with reference at DMSO signal
1
3
the nutrient is out from inside the cell, inhibition of at 2.5 ppm, while the C were 1000-4000 scans
enzyme activity and preventing the synthesis of with the reference of DMSO signal at 39.5 ppm. A
1
nucleid acid and protein .
Bruker VERTEX 70 FT-IR spectrophotometer were
used to record the IR Spectra, KBr discs were used
-1
Organotin(IV)compoundsarewellknownto in all measurements.The range of 4000-400cm was
have wide applications and one of the organometallic applied.Elemental analyses (CHNS) were performed
compounds widely applied and used due to their on Fision EA 1108 series elemental analyser.
5
-21
significants in many biological activities . These
compounds and their derivatives have been known Preparation of organotin (IV) chlorobenzoate
9
-11
12-15
as antifungal , anticancer and antitumour
,
The organotin(IV) chlorobenzoate
16
17-18
20-24
antiviral , antibacterial
biological activity of these compounds because of on the procedure previously reported
the number and basic character of organic ligand procedure was obtained from adaption from the work
attached to metal center Sn . The anions bound to by Szorcsik et al. .For example the procedure in the
Sn metal are the secondary factor in determining the preparation of dibutyltin(IV) dihydroxybenzoate was
and anticorrosion .The compounds used in this work were prepared based
9,14,15,20,21
.These
19
24
organotin (IV) compounds.
as follows:
In this paper, we report the antibacterial
activity of organotin (IV) derivatives of diphenyltin mL methanol was added 0.8 g (0.02 mol) NaOH.The
IV) and triphenyltin (IV) with 3-chlorobenzoic acid reaction mixtures were stirred for about 45 minutes.
againts negative gram bacteria P. aeruginosa and Compound 2 was precipitated out as white solid,
To 3.44 g (0.01 mol) [(C H ) SnCl ] in 50
6 5 2 2
(
positive gram bacteria B. subtilis
filtered off and dried in vacuo till they are ready for
analysis and further reaction.The average yield was
2.33 g (94 %).
EXPERIMENTAL
All reagents used were AR grade.
To 0.4605 g (1.5 mmol) compound 2 in 50
Diphenyltin(IV) dichloride ([(C H ) SnCl ]), mL of methanol was added with 2 mole equivalents
6
5
2
2
triphenyltin(IV) chloride ([(C H ) SnCl]), of 3-chlorobenzoic acid (0.313 g) and was refluxed
6
5 3
3
-chlorobenzoic acid were obtained from Sigma, for 4 hours at 60 – 61°C. After removal of the solvent
sodium hydroxide (NaOH) and methanol (CH OH) by rotary evaporator, the produced compounds
3
were JT Baker products, and the control drug, [(C H ) Sn(OOCC H Cl) ] were dried in vacuo
6
5
2
6
4
2
chloramphenicol were used as received without until they are ready for analysis and further use for
further purification. Negative gram bacteria P. antibacterial activity test. The average yields were
aeruginosa was obtained from Department of more than 90 %.
Microbiology, University of Indonesia, Jakarta and
positive gram bacteriaB. subtilis was obtained from
A similar procedure was also adapted in the
Biochemistry Laboratory, Department of Chemistry, preparation of triphenyltin (IV) derivatives,[(C H ) S
6
5 3
University of Lampung Indonesia.
The UV spectra were obtained using a
n(OOCC H Cl)], where only one mole equivalent of
6 4
the 3- chlorobenzoic acid was used.
UV-Shimadzu UV-245 Spectrophotometer. The Antibacterial Activity Test
measurements were conducted in the UV region Preparation of Media
and were measured in 1 mL quartz-cells. The
The media used for the activity test was
solvent used to dissolve sample was methanol. nutrient agar (NA). 2.8 g of NA was dissolved in 100
The concentration used in the measurements were mL aquadest, heated and sterilized by autoclave at

ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
1135
1
21°C, pressure of 1 atm for 15 minutes. 15 mL of
volumes were then varied. The compound tested
with certained volume was then placed to liquid
NA media, homogenized with vortex and then pour
to petri disc, left them until solidified. The bacteria
suspensions of P. aeruginosa and B. subtilis were
then innoculated on the media at temperature of
37°C for 2-3 days. The growth of bacteria was then
monitored every day.The most effective compounds
tested were a compound which was the compound
with the smallest concentration but the inhibition
steril media was placed on sterilized petri disc.It was
carried out in laminar air flow, and left the media to
solidify.
Antibacterial activity test by diffusion test
One ose ofP. aeruginosa and B. subtilis
was diluted with 2 mL of salin solution (NaCl 0.85%)
and was used as bacteria suspension. 1 mL of the
suspension was then innoculated on NA, flattened
with spreader. 4 paper discs were prepared. The
first paper disc contained the positive control
26
zone was the biggest .
(
chloramphenicol), the second was negative control
RESULTS AND DISCUSSION
containing the solvent used for the test, i.e. DMSO,
the third and fourth paper discs containing the
organotin (IV) compounds tested. All paper discs
were then placed on the surface of media. They
were then incubated for 1 day at 37°C and were
monitored to see the inhibition zone.The compounds
found as the most active, i.e.giving the most effective
inhibition was then tested with the dilution method.
The syntheses of diphenyltin(IV) di
3-chlorobenzoate (2) and triphenyltin(IV)
3-chlorobenzoate (4) were performed by reacting
diphenyltin (IV) dihydroxide (1) and triphenyltin(IV)
hydroxide(3) with 3-chlorobenzoic acid using the a
9, 14,15,21,22
similar procedure reported previously
.
Some important vibration of IR spectra for
the synthesized compound are presented inTable 1.
FromTable 1, compound 1 has characteristic stretch
Antibacterial activity test dilution Test
Based on the result of diffusion test, the
-1
most effective concentration inhibition zones were
obtained for both diphenyltin (IV) di-3-chlorobenzoate
and triphenyltin (IV) chlorobenzoate. These
compounds with the most active concentration
were then dissolved with aquadest- DMSO and the
for Sn-O bond at 693.56 cm .When 1 was converted
-
1
to 2, the new stretch at 1155.86 cm appeared, this
peak is characteristic that Sn-O-C has been formed
which indicated that bond formation between Sn
and carboxyl group from the acid has been formed.
Table 1: Some selected and important IR band of the compounds
ssynthesized
Compound
1
2
3
4
References
Sn-O
Sn-O-C
Sn-ph
OH
C=O
C=C
693.56
-
1077.85
3445.83
-
652.68
1155.86
1072.90
3447.12
1697.63
1591.60
-
-
-
800-600
1158.10
1072.90
3438.79
1629.72
1551.81
1250-1000
1100-1000
3100-3500
1760-1600
1650-1566
1076.83
3615.51
-
1470.87
1479.29
Table2: Microanalytical data of the compounds synthesized
Compounds Elemental Analysis found (Calculated)
C
H
[
[
(C H ) Sn(m-C H (Cl)COO) ] (2)
52.35 (53.42)
58.00 (59.40)
3.05 (3.08)
3.69(3.76)
6
5
2
6
4
2
(C H ) Sn(m-C H (Cl)COO)] (4)
6
5
3
6
4

1136
ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
Table 3.The lmax of the UV spectra of the
to the bound of chlorobenzoate to Sn atom. The
batochromic shift from the starting material to the
synthesized compounds were de to the substitution
of oxygen in 1 and 3 which was replaced by oxygen
organotin(IV) compounds
Compound
lmax (nm)
p→p*
9,14,115,20,21
atom in chlorobenzoate
.
n→p*
1
13
The data of H and C chemical shifts of
the organotin (IV) compounds prepared are tabulated
in Table 4. The characteristic chemical shift in the
spectra of the compounds prepared (Fig.1) obtained
were characterized carefully and compared to some
[
[
(
(C H ) Sn(OH) ] (1)
207.00
235.00
263.00
272.00
6
5
2
2
(C H ) Sn(m-C H
6
5
2
6
4
Cl)COO) ] (2)
2
[
[
(
(C H ) Sn(OH)] (3)
(C H ) Sn(m-C H
204.00
236.00
293.00
285.00
6
5 3
9,14,115,20,21
1
6
5
3
6
4
previous results
. Based on the H NMR
Cl)COO)] (4)
obtained for compound 2 and 4, the chemical shifts
of phenyl protons attached to tin metal as expected
appeared in the range of 7.4 – 7.6 ppm, while the
protons in the benzoate ring were at 7.7-7.9 ppm.The
chemical shifts of carbon for both compounds also
-
1
The other characteristic vibration is at 1697.63 cm ,
specific for C=O stretch, indicated that carbonyl is
present in the compounds.
9,14,115,20,21
close to the reported values
. The carbon
Table 2 presents the data of microanalysis
for the organotin (IV) compounds synthesized.
The values obtained generally are very good and
compared to the calculated theoretical values were
less than 1%.
in the carboxyl group as predicted appeared in the
region of 165-166 ppm, while carbon of the phenyl
ligand appeared in d of 128-130 ppm. The carbons
in the chlorobenzoate as expected appeared in
d range of 130 – 135 ppm close to the reported values
The lmax. of all compounds were obtained of similar compounds9,14,115,20,21
.
by UV spectroscopy analyses.The data obtained are
shown inTable 3.Although the shift was not very high,
but It is clear that there was some important shifting
change in the lmax for each compound.In the spectra,
all compounds produced 2 peaks due to transition
of p→p and n→p* . For example in compounds 1,
the lmax observed were 207 and 263 nm. In 3 and
The results of antibacterial by diffusion
method are shown Table 5. The increase of
halozone diameter was observed with the increase
of concentration of all compounds tested. The
triphenyltin (IV) 3-chlorobenzoate was more active
in inhibiting B. subtilis than P. Aeruginosa, as the
4, there were large shift in the p→p* transition, due
Table 4: H and 13C spectra of the organotin(IV) compounds
1
Compound
H in phenyl
ppm)
H in benzoate
(ppm)
C in phenyl
and benzoate (ppm)
(
[
(C H ) Sn(m-
H2 & H6 7.538
(d,4H); H3 &H5
7.703-7.991 (m)
C1-6 (phenyl): 129,3 –
128,8; C7 166,9; C8
131,6; C9 130,3; C10
134,2; C11 1340 ; C12
6
5 2
C H (Cl)COO) ]
6
4
2
7.554 (t, 4H); H4
7.570 (t,2H)
130,0; C13 128,4
[
(C H ) Sn(m-
H2&H6 7.46
(d,6H); 3&H5 7.43
7.84-7.85 (d)
C1-6 (phenyl): 129.3 -
128.8
6
5 3
C H (Cl)COO)]
6
4
(t 6); H4 7.41
128.8; C7: 165; C8:
1
31.6; C9: 130.3; C10:
34.2; C11: 134.0;
C12: 130.0; C13: 128.4
1

ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
1137
Cl
inhibition zone observed was bigger in B. subtilis,
although it was still smaller than the inhibition zone
of the positive control used, chloramphenicol. The
differnces in inhibition zone of both bacteria is
because B. subtilis has higher sensitivty compared
3
12
4
1
3
O
C
11
10
2
5
O
O
7
8
1
6
9
Sn
C
O
to P. Aeruginosa perhaps due to the differences of
25-27
cell wall structure of the bacteria
.
Based on data presented in Table 6, the
(a)
-
4
compound 4 at concentration of 3.956 x 10 M,
was the most effective concentration that inhibit the
growth of the both bacteria. The diffusion method
has been applied in order to find the smallest
concentration that is able to inhibit the growth of
bacteria, thus the ratio comparison has been used
to determine the effectivity of the two compounds
tested, by observing the wide of clear zone per the
number of active compound in the media.The result
2
1
3
4
Cl
6
1
3
5
12
11
Sn
7
C
O
8
O
9
10
-
4
showed that the concentration of 6 at 3.956 x 10
(b)
M (200 ppm) has the biggest ratio, even when it is
compared with the increased of the concentration of
the compound tested.
Fig. 1: Structure of (a) diphenyltin(IV) di-
-chlorobenzoate, (b) triphenytin(IV) 3-
chlorobenzoate.
3
Table 5: The result of diffusion test againts P. aeruginosa and B. subtilis
Compound
Inhibition Zone (cm)
2
00
250
300
400
500
(
ppm)
(ppm)
(ppm)
(ppm)
(
ppm)
P.a B.s
P.a B.s
P.a B.s P.a B.s
P.a
B.s
[
[
[
[
[
C H (Cl)COO)]
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
4
(C H ) Sn(O)] (1)
0.8
-
0.8
0.9
-
0.9
0.9
-
1.0
1.0
-
1.1
1.0
-
1.1
1.4
2.5
6
5 2
(C H ) Sn(m-C H (Cl)COO) ](2)
0.8
-
0.8 1.2
1.5 2.5
0.8
-
0.9 1.3
1.5 2.5
6
5
2
6
4
2
(C H ) Sn(OH)] (3)
-
6
5 3
(C H ) Sn(m-C H (Cl)COO)] (4)
0.8 1.0
1.5 2.5
0.7 1.1
1.5 2.5
0.9
1.5
6
5
3
6
4
Chloramphenicol
P.a. = Pseudomonas aeruginosa, B.s. = Bacillus subtilis,
Positive control = Chloramphenicol
Table 6:The inhibition percentage of ccompound tested against
P. aeruginosa and B. subtilis
Compound
% Inhibition
2
00
ppm)
B.s
250
(ppm)
300
400
(ppm)
500
(
(ppm)
(ppm)
P.a
P.a
B.s
P.a
B.s
P.a
B.s
P.a
B.s
[
(
(C H ) Sn(m-C H
-
-
-
-
0.003
-
0.003
-
-
-
6
5
2
6
4
Cl)COO) ] (2)
2
[
(
(C H ) Sn(m-C H
Cl)COO)] (4)
0.004 0.005 0.003 0.004 0.003 0.004 0.002 0.003 0.002 0.003
6
5
3
6
4
P.a. = Pseudomonas aeruginosa, B.s. = Bacillus subtilis,
Positive control = Chloramphenicol

1138
ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
Table 7:The result of dilution test of comppound 4 against P. aeruginosa and B. subtilis
Bacteria
Volume of 4 (mL)
0
.5
1
1.5
++++++
2
2.5
—
P. aeruginosaB. subtilis
++++++++++ ++++++++
++++
Note of Bacterial growth:
+
+
+
++++
+++
++
= very high
= high
= medium
= little
+
+
+
-
= very little
= no grwoth
The result of dilution test is shown in triphenyltin(IV) 3-chlorobenzoate were prepared.
Table 7. The compound 4 in concentration 3.956 Based on the result obtained, they have shown to
-
4
x 10 M (200 ppm) with volume used 2.5 mL per be potentially used as antibacterial substances.
media was able to inhibit maximally the growth of The triphenyltin(IV) derivative has been shown to
both bacteria , while at other volumes, the bacteria be more active than diphenyltin(IV) derivative. This
were still able to grow.
finding was in line with other data relating to the
number of carbon atom present in the compound
The microorganism inhibition mechanism and might also relate to the ability of phenyl ligand
by antibacterial substances may be caused by to draw electron from the metal center as a result
some factors, such as the disturbance in the the metal became more positive and reacted actively
compound composition of cell wall, the increase of with electronegative cell of bacteria, thus the growth
cell membrane permeability which cause the loss of of bacteria was disrupted. However, we aim to have
component cell structure, the inactivation of enzyme a better antibacterial substance which has much
and destruction or damaging the function of genetic smaller inhibition concentration.
2
5
materials .
ACKNOWLEDGMENT
In this biologial activity test, the compound
where its compound is more electropositive than 3,
4
Annisa would like to thank to the Head of
disturb the electronegative bacteria cell wall, thus the Sekolah Tinggi Ilmu Kesehatan Faletehan Serang-
interaction cause the disruption of bacteria growth.
Banten for financial support for her study especially
for the final research.Thanks also go to Prof. Bohari
M.Yamin, Universiti Kebangsaan Malaysia for helping
in doing microanalysis and Prof. Dr. Hasnah Osman
CONCLUSION
The derivatives of organotin(IV) of School of Chemistry, University of Sains Malaysia
compound, diphenyltin(IV) di-3-chlorobenzoate and for NMR experimentation.
REFERENCES
1
2
.
.
Jawetz, E. and M. Adelberg. Mikrobiologi
Kedokteran Edisi Ke-3.Alih Bahasa :Huriwati
Hartanto dkk. Penerbit Buku Kedokteran
ECG.Jakarta. 2005.
Nursal, S.Wulandari, W.S.Juwita. Bioaktifitas
Ekstrak Jahe (Zingiber officinale Roxb.) dalam
Menghambat Pertumbuhan Koloni Bakteri
Escherichia coli dan Bacillus subtilis. J.
Biogen. 2006, 2, 64-66. (in Indonesian)
Irianto, K.. Mikrobiologi Menguak Dunia
Mikroorganisme Jilid 2. Bandung: CV.Yrama
Widya. 2007 (In Indonesian)
3.

ANNISSA et al., Orient. J. Chem., Vol. 33(3), 1133-1139 (2017)
1139
4
.
Pelczar,M.J, Chan, E.C.S.Dasar-
dasarmikrobiologi2.Diterjemahkan oleh
Hadioetomo RS, Imas T, Tjitrosomo SS,
Angka SL. PenerbitUniversitas Indonesia.
Jakarta. 1986; pp.489-522
18.
Win Y. F., Teoh, S. G., Vikneswaran, M. R.,
Sivasothy,Y., Ha,S. T., Ibrahim, P. Aus. J.
Basic Appl. Sci. 2010,4,: 5923-5931
Pellerito, L.; Nagy, L., Coord. Chem. Rev.
2002, 224, 111-150.
Kurniasiah, H , Nurissalam, M ,Iswantoro, B
, Afriyani,H , Qudus,H.I., Hadi., S.,Orient. J.
Chem. 2015, 31, 2377-2383.
Hadi, S., Rilyanti, M.; Nurhasanah.
ModernAppl. Sci. 2009, 3, 12-17.
Rastogi, R.B.; Singh, M.M.; Singh, K.;
Yadav,M. Port. Electrochim. Acta. 2005, 22,
315-332.
19.
20.
5
6
7
8
.
.
.
.
Kang, W., Wu, X., Huang, J.J.Organo.Chem.
2009, 694, 2402-2408.
Win,Y.F., Teoh,S.G., Lim, E.K., Ng,S.L., Fun,
H.K. J. Chem. Crystal. 2008, 38, 345-350.
Alama, A., Tasso, B. , Novelli, F., Sparatore,
F.,Drug Discov. Today 2009,14, 500-508.
Affan, M.A., Foo,S.W., Jusoh,I., Hanapi, S.,
Tiekink, E.R.T. Inor. Chem. Acta 2009,362,
21.
22.
5
031-5037.
Hadi, S.; Irawan, B.; Efri. J. Appl. Sci. Res.
008, 4, 1521-1525.
9
1
1
1
1
1
1
1
.
23.
24.
Singh, R.; Chaudary, P.; Khausik, N.K. Rev.
Inorg. Chem. 2010, 30, 275-294.
2
0.
1.
2.
3.
4.
5.
6.
Manav, N., Ghandhi, N., Kaushik,N.K. J.
Therm. Anal. Calorom. 2000,61: 127-134.
Singh, R., Kaushik, N.K. Spec. Acta Part A:
Mol. Biomol. Spectr.2008,71, 669-675.
Mohan, M., Agarwal, A., Jha, N.K. J. Inor.
Biochem. 1988,34, 41-54.
Szorcsik, A.; Nagy, L.; Gadja-Schrantz,
K.;Pellerito, L.; Nagy, E.; Edelmann, E.T.
J.Radioanal. Nucl. Chem. 2002, 252, 523-
530.
Gielen, M.J.Braz.Chem.Soc.2003, 14, 870-
25.
Jawetz, E., Melnick, L.J.; Adelberg, A.E..
Mikrobiologi untuk Profesi Kesehatan ed 16,
terjemahan Tonang, H.egc Penerbit Buku
Kedokteran. Jakarta. 1986; Hal.31,34,145-
147,150-152.
Lorian, V. Antibiotics in Laboratory Medical.
Wiliam and Wilkins Co.Baltimore. London.
1980; Hal. 1-22, 170-178, 511-512.
Ibrahim, M.Mikrobiologi: Prinsip dan Aplikasi.
Surabaya: Unesa University Press. 2007. (In
Indonesian)
8
77.
Hadi, S.;Rilyanti, M.;Suharso.Indo.J.Chem.
012, 12, 172-177.
Hadi, S.;Rilyanti, M.Orient.J.Chem.2010,26,
75-779.
2
7
26.
27.
Singh, N.K., Srivastava, A., Sodhi, A.,
Ranjan, P. Transit. Metal Chem. 2000, 25,
133-140.
1
7.
Maiti, A., Guha, A.K., Ghosh, S. J. Inor.
Biochem. 1988,33, 57-65.