Carbon 39 (2001) 19631969

Antibacterial activated carbons prepared from pitch containing

organometallics
Hisashi Tamai a , Nobuyoshi Katsu a , Kazuhisa Ono b , Hajime Yasuda a , *
a

Department of Applied Chemistry, Faculty of Engineering, Hiroshima University, Higashi-hiroshima 739 -8527, Japan
Department of Fermentation Technology, Faculty of Engineering, Hiroshima University, 1 -4 -1 Higashi-hiroshima 739 -8527, Japan
Received 19 July 2000; accepted 29 December 2000

Abstract
A simple method for the preparation of antibacterial activated carbons was reported. The antibacterial activated carbons
uniformly contain metal oxides, i.e. CaO, MgO, NiO, CoO and ZnO. These activated carbons were readily prepared by
steam activation of pitch containing organometallics, i.e. CaI 2 , t-BuMgBr, Ni(acac) 2 , Co(acac) 3 , and Zn(acac) 2 . The
antibacterial activities for B. subtilis, S. aureus, and E. coli were estimated by the growing inhibitory effect in the solutions
containing activated carbons and by haloes at surrounding of activated carbons on agar. The CaO, MgO, CoO, and ZnO
dispersed activated carbons exhibited obviously the antibacterial activity for B. subtilis both in the solutions and on agar. The
MgO and CoO dispersed ones exhibited a growing inhibition effect for S. aureus. On the other hand, no inhibitory effect was
observed for the growing of E. coli. 2001 Elsevier Science Ltd. All rights reserved.
Keywords: A. Activated carbon, pitch; B. Activation

1. Introduction
Activated carbons have attracted much attention as one
of the effective adsorbents. For wide applications of
activated carbons as adsorbents, the controlling of pore
size and its distribution is important. Until a recently there
have been many investigations on the controlling of pore
size. Recently, we have developed highly mesoporous
carbons by activating pitch containing rare earth metal
complexes with steam [13], and then we showed that
metal or metal oxide particles were uniformly dispersed in
activated carbons obtained from pitch containing organometallics in terms of electron probe microanalysis
(EPMA) and X-ray diffraction (XRD) analyses [3,4].
On the other hand, the antibacterial activity of activated
carbons are also very important for application in water
purification systems, e.g. purification of drinking water
because the growing of bacteria on the surface of activated
carbons spoils the function of activated carbon and resulting water may be polluted by bacteria. In general, the
antibacterial activities of activated carbons are available as
*Corresponding author. Tel.: 181-824-24-7731; fax: 181824-22-7191.
E-mail address: yasuda@hiroshima-u.ac.jp (H. Yasuda).

Ag deposited activated carbons prepared by impregnation

methods [57]. However, in the case of Ag supported
activated carbons prepared by impregnation or deposition
methods, there arise some problems, e.g. needing a few
steps for Ag-supporting such as impregnation in Ag
solutions and heat-treatment of activated carbons, a decrease in surface area and pore size by deposited Ag, and
rapid elution of Ag ions at the initial stage of usage [5].
Sawai et al. [810] reported that some metal oxides
powders e.g., MgO, CaO, and ZnO showed a growth
inhibitory effect for bacteria such as Staphylococcus
aureus (S. aureus) and Bacillus subtilis (B. subtilis). Based
on these results., we expected that metal oxides particles
uniformly dispersed in activated carbons should show
antibacterial activity. Especially, finely and uniformly
dispersing of metal oxide particles in activated carbons
plays an important role to realize the excellent antibacterial
effect.
From these points of view, we attempted the preparation
of the antibacterial activated carbons without complicated
steps used for the preparation of Ag-supporting activated
carbon. We supposed the formation of metal oxides
simultaneously with the formation of pores in activated
carbons by activating pitch containing organometallics.
The obtained activated carbons exhibited antibacterial

0008-6223 / 01 / $ see front matter 2001 Elsevier Science Ltd. All rights reserved.
PII: S0008-6223( 01 )00003-3

1964

H. Tamai et al. / Carbon 39 (2001) 1963 1969

activities for B. subtilis and S. aureus, although their

activities were lower than that of Ag deposited activated
carbons. We show that the antibacterial activated carbons
can be prepared by the simple procedure.

2. Experimental

2.1. Materials
Pitch was coal tar pitch from Osaka gas Co. (softening
point: 72.28C, C: 93.2%, H: 4.7%, N: 1.1%). CaI 2 , tBuMgBr, Ni(acac) 3 , Fe(acac) 3 , Co(acac) 3 , Al(acac) 3 , and
Zn(acac) 2 were purchased from Kanto Chemical and used
without further purification. Manganocene (Cp 2 Mn) was
synthesized as follows [11]: cyclopentadiene (1.59 mmol)
was added slowly to sodium shot (3.65 g) in THF (100
ml), and then to this solution manganous chloride (10 g)
was added. The mixture was refluxed for 3 h and THF was
removed under vacuum. The resulting Cp 2 Mn was sublimed in vacuo. Dark brown Cp 2 Mn crystals were obtained
(yield; 2 g). Tetrahydrofuran (THF) (Kanto Chemical) was
dried over Na / K alloy and distilled before use. S. aureus
(HUT005), B. subtilis (IFO3134), and Escheritia coli (E.
coli) (HUT8106) were obtained from Fermentation Technology Department of Hiroshima University.

2.2. Carbons
Various activated carbons were prepared by steam
activation of pitch containing organometallics. Pitches
containing organometallics were obtained by mixing a
THF solution of pitch with THF solutions of organometallics. A typical preparation method is as follows: 4.79 g of
pitch was placed in a 300 ml two necked round bottom
flask and volatile components were removed under reduced
pressure at 1208C. The pitch was dissolved in 90 ml of dry
THF. Zn(acac) 2 (1.209 g) dissolved in 90 ml of dry THF
was added to the pitch solution. After stirring for 2 h, THF
was removed by flash distillation.
Activated carbons were obtained by activation of pitch
containing organometallics with steam at 9008C. Ag
supported activated carbons were prepared by the impregnation method [6]. Activated carbon prepared from
organometallics-free pitch by steam activation was impregnated in aq. solutions of AgNO 3 for 24 h. After
evaporation of AgNO 3 solution, the activated carbon was
heat-treated at 4208C for 1 h.

2.3. Methods
Specific surface area and pore size distribution were
determined by BET method, and t-plots (for micropore)
and BJH [12] (for mesopore) methods, respectively, by
applying N 2 adsorption isotherms measured using a Quantachrome Autosorb-6. XRD analyses were performed using

a Rigaku RDA-1B system with Cu Ka radiation. The

dispersion of metal oxides in activated carbons was
observed by electron probe microanalysis (EPMA) with a
JEOL JXA-8900. The metal contents in carbons were
measured by particle-induced X-ray emission (PIXE)
analysis and ash contents in activated carbons. PIXE
analysis was performed using a van de Graaff accelerator
of Hiroshima University. The intensity of the characteristic
X-ray generated by irradiation of H 1 was measured.
We estimated antibacterial activities of metal oxides
particles dispersed in activated carbons by the growing
inhibitory effect for B. subtilis, S. aureus, and E. coli. A
growing inhibitory effect was determined by two kinds of
methods; (1) measurement of bacteria growing in the
solutions containing activated carbons as follows: a portion
of B. subtilis was inoculated in 5 ml of sporulation
medium (beef extract 10 g, poly-pepton 10 g, NaCl 5 g / 1 l
of pure water). The medium was incubated at 378C for 18
h. The spores were inoculated in 5 ml of sporulation
medium and to this medium activated carbons were added.
The spores were filtered after the prescribed time. The
filtrate (0.5 ml) was diluted by 4.5 ml of sporulation
medium. The bacteria growing was determined by measuring optical density of the diluted solution. Optical density
(OD 660 ( l 5660 nm)) was measured by UVVisible
spectrometer(Pharamacia LKB Biochrome 4060 UVVisible Spectrometer).
(2) Measurement by the halo method as follows [12]:
bacteria was inoculated in 5 ml of a sporulation medium
(poly-pepton 10 g, yeast extract 2 g, and magnesium
sulfate 1 g / 1 l of pure water), and this solution was
incubated at 378C for 18 h. The spores were inoculated in
100 ml of sporulation medium and incubated at 378C for
18 h. Agar containing these spores was placed onto agar
plates containing activated carbons and dried for a few
minutes. After incubating for 24 h at 378C, the lengths of
haloes formed surrounding the activated carbons were
measured.

3. Results and discussion

Table 1 shows the carbonized yields and pore characteristics of the activated carbons obtained from pitches
containing various organometallics. The addition of CaI 2
lowered the yield and BET specific surface area. The
formed CaO accelerated the consumption of carbon precursors such as pitch during steam activation and as a
result, the formed pores were supposed to be destroyed
throughout activation. On the other hand, the addition of
other organometallics, i.e. t-BuMgBr, Zn(acac) 2 , Ni(acac) 3 ,
Fe(acac) 3 , Co(acac) 3 , and Al(acac) 3 into pitch promotes the
formation of pores and BET specific surface areas are
higher than that from organometallics-free pitch in spite of
the short activation time. Especially, the promoting effect
for pore formation is remarkable in the addition of

H. Tamai et al. / Carbon 39 (2001) 1963 1969

1965

Table 1
Preparation of activated carbons from pitches containing various organometallics a
Sample

Organometallics

Metal
in pitch
(wt.%)

Activation
time
(min)

Yield
(%)

BET surface
area
(m 2 / g)

Mean pore
size
(nm)

CaAC
MgAC
ZnAC
MnAC
NiAC
FeAC
CoAC
AlAC
AC

CaI 2
t-BuMgBr
Zn(acac)2
Cp 2 Mn
Ni(acac) 2
Fe(acac) 3
Co(acac) 3
Al(acac) 3

5.0
5.0
5.0
2.5
5.0
5.0
5.0
5.0

3
15
15
25
12
15
18
18
30

14.8
40.2
26.3
31.8
28.2
36.9
24.6
32.0
24.7

0.4
118
150
76
131
159
159
221
88

50.0
3.1
2.6
3.6
3.2
3.8
3.8
2.2
2.5

Activation temp.: 9008C.

Co(acac) 3 and Al(acac) 3 . Fig. 1 shows the pore distributions of activated carbons obtained from pitches containing
t-BuMgBr and Zn(acac) 2 . Micropores of about 0.5 nm of
pore radius were mainly developed in activated carbon
obtained from pitch containing t-BuMgBr, whereas pores
of large size (mesopores) were formed in addition to
micropores by the addition of Zn(acac) 2 . Similarly, a large
ratio of micropores and small ratio of mesopores were
formed in activated carbons obtained from pitches containing other organometallics.
The decomposition of organometallics results in the
formation of metal or metal oxide in the course of
activation of pitch [2,4]. We measured metal contents in
the activated carbons obtained. The results are shown in
Table 2. Metal contents in the activated carbons range
from 4.3 to 9.8 wt.% in the cases of Zn, Ni, Fe, Co, and
Ni. These values were lower compared with the theoretical
values calculated from metal contents in pitch (5 wt.%). A
part of metal atoms is supposed to disappear throughout
the decomposition of organometallics during activation.
On the other hand, the detection of light atoms such as Ca

and Mg by PIXE analysis was impossible. Based on metal

contents determined from ash contents in activated carbons
obtained from pitches containing CaI 2 and t-BuMgBr, it is
supposed that most metal atoms remain in the activated
carbons. In addition, XRD analysis indicated the formation
of crystals of CaO and MgO in the activated carbons as
described below.
Sawai et al. [810] reported that some metal oxides
showed antibacterial activities. We analyzed metal species
formed from organometallics in activated carbons during
activation by XRD analyses. Fig. 2 shows XRD patterns of
the activated carbons obtained from pitches containing
CaI 2, t-BuMgBr, Zn(acac) 2 , and Cp 2 Mn. The peaks due to
CaO, MgO, and ZnO on each XRD patterns were observed
in addition to the broad peak of carbon crystallites. As
shown in Fig. 2, MnO and Mn 2 O 3 particles were formed
from Cp 2 Mn. Similarly, the formation of metal oxides, i.e.
Al 2 O 3 , and Fe 2 O 3 from Al(acac) 3 and Fe(acac) 3 , respectively, was observed. On the other hand, the formation of
metal particles, i.e. Co and Ni in addition to metal oxides
such as CoO and NiO from Co(acac) 3 and Ni(acac) 3 , was
observed.
Table 2
Metal content in activated carbons obtained from pitches containing organometallics

Fig. 1. Pore distributions of activated carbons obtained from

pitches containing t-BuMgBr and Zn(acac) 2 (s); t-BuMgBr, (d);
Zn(acac) 2 .

Sample

Metal
in pitch
(wt.%)

Metal
in carbon
(wt.%)

CaAC
MgAC
ZnAC
MnAC
NiAC
FeAC
CoAC
AlAC

5.0
5.0
5.0
2.5
5.0
5.0
5.0
5.0

25.0 a
9.9 a
4.4 b
0.8 b
9.8 b
6.5 b
8.6 b
4.3 b

a
b

Determined from ash content.

Determined by PIXE.

1966

H. Tamai et al. / Carbon 39 (2001) 1963 1969

Fig. 2. XRD patterns of activated carbons obtained from pitches

containing organometallics (a), CaI 2 ; (b), t-BuMgBr; (c),
Zn(acac) 2 ; (d), Cp 2 Mn.

Fig. 4. Optical densities of sporulation medium containing Ca

AC for B. subtilis. Concentration of activated carbon (d); CaAC
10 mg / ml, (m); CaAC 30 mg / ml, (n); AC 30 mg / ml, (s);
none.

The dispersion states of metal oxides in the activated

carbons obtained from pitches containing organometallics
are important for antibacterial activity of activated carbons.
The homogeneous dispersion of metal oxides were observed by EPMA. SEM and EPMA images of the activated
carbon obtained from pitch containing Zn(acac) 2 are
exemplified in Fig. 3. ZnO were dispersed almost uniformly in activated carbon. The distributions of other metal
oxides were also uniform as observed for ZnO.
The antibacterial activities for B. subtilis, S. aureus, and

E. coli were investigated. Bacteria were incubated in

sporulation medium containing activated carbons and the
change of optical density (OD) of medium with incubation
time was measured. Figs. 47 show the results on the
activated carbons obtained from pitches containing CaI 2 ,
t-BuMgBr, Zn(acac) 2 , and Cp 2 Mn, for B. subtilis. As
shown in Figs. 4 and 5, in the cases of the activated
carbons obtained from pitches containing CaI 2 and tBuMgBr, no increase of OD with time was observed while

Fig. 3. EPMA image of activated carbon obtained from pitch containing Zn(acac) 2 .

H. Tamai et al. / Carbon 39 (2001) 1963 1969

Fig. 5. Optical densities of sporulation medium containing Ma

AC for B. subtilis. Concentration of activated carbon (d); Mg
AC 10 mg / ml, (m); MgAC 30 mg / ml, (j); MgAC 50 mg / ml,
(n); AC 30 mg / ml, (s); none.

the OD of medium without activated carbon or the OD of

medium containing activated carbon obtained from organometallics-free pitch increased with incubation time.
These results obviously indicate that the activated carbons
obtained from pitches containing CaI 2 and t-BuMgBr
possess strong antibacterial activities for B. subtilis. In
addition, the activated carbons obtained from pitches
containing Co(acac) 3 and Ni(acac) 3 showed the same
antibacterial effect for B. subtilis as that from pitch
containing t-BuMgBr. As shown in Fig. 6, the OD of
sporulation medium for activated carbon containing ZnO
slightly increases at the initial stage of incubation and the
value of OD decreased with increasing the amount of

Fig. 6. Optical densities of sporulation medium containing Zn

AC for B. subtilis. Concentration of activated carbon (m); ZnAC
30 mg / ml, (d); ZnAC 50 mg / ml, (n); AC 30 mg / ml, (s);
none.

1967

Fig. 7. Optical densities of sporulation medium containing Mn

AC for B. subtilis. Concentration of activated carbon (d); Mn
AC 30 mg / ml, (m); MnAC 50 mg / ml, (n); AC 30 mg / ml, (s);
none.

activated carbon added in incubation medium. The ZnO

dispersed activated carbon is supposed to suppress growing
of bacteria, although the antibacterial activity was lower
than that of the MgO dispersed one. On the other hand, as
shown in Fig. 7, the activated carbon containing MnO and
Mn 2 O 3 exhibited no growing inhibitory effect for B.
subtilis.
Fig. 8 shows the changes of ODs of sporulation medium
containing activated carbons for S. aureus with incubation
time. Mediums containing activated carbons prepared from
pitch containing Zn(acac) 2 or metal oxide-free activated
carbon exhibited the same growing curves as that in
medium without activated carbon. There is no antibacterial
activity in activated carbon containing ZnO for S. aureus.

Fig. 8. Optical densities of sporulation medium containing 30

mg / ml of activated carbons for S. aureus (j); CaAC, (s);
MgAC, (n); ZnAC, (h); NiAC, (m); AC, (d); none.

H. Tamai et al. / Carbon 39 (2001) 1963 1969

1968

Fig. 9. Optical densities of sporulation medium containing 30

mg / ml of activated carbons for E. coli (j); CaAC, (s); Mg
AC, (n); ZnAC, (h); NiAC, (); CoAC, (m); AC, (d);
none.

On the other hand, the increase in ODs of mediums

containing the NiO, MgO, and CaO dispersed activated
carbons is low, compared with the OD of medium containing metal oxide-free activated carbon. These activated
carbons exhibited an inhibitory effect for the growing of S.
aureus.
On the antibacterial activity for E. coli, Fig. 9 shows the
relation between ODs of sporulation mediums containing
activated carbons and incubation time. ODs of mediums
containing metal oxides dispersed activated carbons
scarcely increased with time. However, the OD of medium
containing metal oxide-free activated carbons was also
low. The growing inhibitory effect of the metal oxides
dispersed activated carbons for E. coli seems due to the
adsorption of E. coli on activated carbons rather than the
antibacterial activity of metal oxides.
Furthermore, in order to clarify the antibacterial activity
of metal oxides dispersed activated carbons, we measured
the growing inhibitory effect on agar by the halo method.
Table 3 shows the lengths of haloes formed surrounding
the activated carbons. The activated carbons containing

Table 3
Growth inhibitory effect for bacteria on agar
Sample

CaAC
MgAC
ZnAC
NiAc
FeAC
CoAC
AlAC
ACAg5
ACAg10

CaO, MgO, ZnO, and CoO show the formation of haloes

of almost the same lengths for B. subtilis. This formation
of haloes indicates that these activated carbons give the
inhibitory effect for the growth of B. subtilis. However, the
lengths of haloes are half of that by Ag supported activated
carbons, which were prepared by the impregnation method
using aq. solution of AgNO 3 . On the other hand, NiO
dispersed activated carbon for B. subtilis and S. aureus and
CaO dispersed activated carbon for S. aureus, no growing
inhibitory effect was observed on agar in spite of the fact
that it exhibited the antibacterial activity in the solutions.
This may be due to the difference in contact area of
activated carbons to bacteria. The contact area of activated
carbon in solutions is supposed to be larger than that on
agar. These results suggest that the antibacterial activity of
NiO dispersed activated carbon is not so strong as that of
MgO dispersed activated carbon.The activated carbons
obtained from pitches containing organometallics showed
no inhibitory effect on agar for E. coli, in spite of that Ag
supported activated carbon exhibited strong inhibitory
effect.

4. Conclusion
The antibacterial activated carbons can be simply prepared by steam activation of pitch containing organometallics. Metal oxides were formed uniformly in activated
carbons and the antibacterial activities were dependent on
the kind of metal oxides. The CaO, MgO, CoO, and ZnO
dispersed activated carbons obtained from pitches containing CaI 2 , t-BuMgBr, Co(acac) 3 , Ni(acac) 2 , and
Zn(acac) 3 , respectively, exhibited good antibacterial activities for B. subtilis. The MgO and CoO dispersed ones
presented a growing inhibitory effect in sporulation solutions and on agar for S. aureus. On the other hand, no
inhibitory effect was observed for the growing of E. coli.
The antibacterial activated carbons do not need the complicated procedures such as that for the preparation of Agsupporting activated carbon.

Acknowledgements

Halo length
(mm)
B. subtilis

E. coli

S. aureus

4.2
4.0
4.2
0
0
4.5
0
9.9
9.9

0
0
0
0
0
0
0
4.0
6.0

0
8.5
0
0
0
6.5
0
9.0
13.3

This work is partially supported by a Grant-in-aid for

Research for the Future Program, Nano-carbon, from the
Japan Society for the Promotion of Science.

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