Article

pubs.acs.org/IECR

Impact of Mixed Inhibitor on Electrochemical Behavior of Inland

Water Biofilm Formed on 316L Stainless Steel
Dharmarajan Sridharan, Chandrasekaran Karthikeyan, Sundaram Maruthamuthu, Subbiah Manoharan,
and Narayanan Palaniswamy*
Corrosion and Material Protection Division, CSIR-Central Electrochemical Research Institute, Karaikudi 630 006, Tamil nadu, India

ABSTRACT: The electrochemical behavior of biofilm on AISI 316L stainless steel was studied in the mixed inhibitor system:
amino trimethylene phosphonic acid with zinc sulfate (ATMP + Zn2+) added inland water system. The natural biofilm shifts
potential toward positive side, about 380 mV vs SCE and in the presence of inhibitor, the biofilm does not shift the potential
toward positive side. Cathodic polorization and cyclic voltammogram explained the presence of manganese oxide and peroxide in
the natural biofilm. There were no oxidation and reduction peak in the inhibitor treated (industrial) biofilm because of removal
of cations in the biofilm by inhibitor. The natural biofilm has thick matrix with extracellular polysaccharides (EPS) on 316L SS. In
the presence of inhibitor in the water, distinct rod shaped cells were noticed without any EPS. The inhibitor added system
showed the lowest passive current (ip) and high resistance (Rct) when compared to the natural biofilm.

1. INTRODUCTION Scheme 1. Molecular Structure of ATMP

Metallic surfaces exposed to different aqueous media adsorb
organic/inorganic matter from the surrounding environment.
Bacteria, microalgae, fungi, and protozoans get attached to the
materials within a day where their extracellular metabolites
cause the formation of an extremely complex micro layer called
biofilm. The natural biofilm consists of aerobic and anaerobic
bacteria viz., manganese oxidizers, acid producers, hetero-
trophic bacteria, iron oxidizers, sulfate reducing bacteria, etc.
The role of the natural biofilm formed on passive alloy is an
interesting aspect of research on microbial influenced corrosion
(MIC) that has been studied by many investigators1−8 who
proposed many mechanisms. The role of manganese oxidizing
bacteria on the stainless steel is significant.7,8 It interacts with
the oxide film (n/p-type) semiconductors on stainless steel and
determines its electrochemical behavior. Extracellular poly-
saccharide (EPS) is an important factor for the creation of
electrochemical cell on passive alloys.9 There is no report
available on the role of biofilm on passive alloys in the inhibitor
treated system in literature. Inhibitors are also added in cooling
water system to combat corrosion, where the biofilm consists of
various types of bacteria and can be named as industrial biofilm.
Many industrial processes use the amino trimethylene
phosphonic acid (ATMP) to prevent scale formation10 and
corrosion in cooling water system. It is assumed that the
inhibitor/biocide resistant strains in industry may create
different consortia, which may influence on the electrochemical
behavior of stainless alloys. Nobody has investigated the role of
biofilm in passive alloys while adding inhibitor in cooling water
industry. In the present study, a mixed inhibitor of ATMP
(Scheme 1) with ZnSO4 system was used to form the industrial
biofilm on AISI 316L SS and the electrochemical behavior of
the industrial biofilm formed in pond water was investigated.
2. MATERIAL AND METHODS
2.1. Specimen Preparation. The composition of AISI
316L stainless steel employed is given in Table 1. Specimen of
dimensions 5 × 2.5 × 0.2 cm in size was used for
electrochemical studies. An electrical lead of nichrome wire
was spot-welded at the top end of each specimen. The
specimens were ground with SiC emery paper down to 1000
grit and then treated in a process of sonication with acetone for
5 min to remove any residual oil-based contamination. The
specimens were kept for 3−4 min in the ultrasonication process
with Milli-Q water. 316L SS specimens were treated by the
passivation process with 10% nitric acid for 20 min at 60 °C
and the passivated 316L SS specimens were used for all
experimental purposes.
2.2. Biofilm Formation. The freshwater was collected from
Ammon pond,11 Karaikudi, India, and the physio-chemical
characteristics of the water were done as per the methods
described by APHA.12,13 The ATMP inhibitor used in the
present study was supplied by Solutia Co. as an aqueous
solution (50% w/w). Analytical grade ZnSO4 was obtained
from Fisher Scientific Inc. The stock solutions of inhibitors
were made by diluting 0.01 g L−1 of ATMP and dissolving
0.005 g L−1 of ZnSO4. The specimens were exposed to
inhibitor (ATMP + Zn2+) treated natural fresh water and
natural water (without inhibitor) to form a biofilm. The natural
freshwater was sterilized in an autoclave at 121 °C, and further
Received: April 23, 2013
Revised: October 13, 2013
Accepted: October 21, 2013
Published: October 21, 2013

© 2013 American Chemical Society 16175 dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181
Industrial & Engineering Chemistry Research Article

Table 1. Chemical Composition of 316L Stainless Steel

C Mn P S Si Cr Ni Mo Fe
mass (%) 0.03 1.98 0.034 0.02 0.84 17.85 12.11 2.12 balance

SS specimens were immersed in sterilized freshwater as control using Gamry Potentiostat (EIS 300) with lock-in-amplifier
system. The immersion step was done carefully, so that the model (SRB10). The impedance test was carried out at steady
water line on the sample was sufficiently below the welded state of OCP with an amplitude of 5 mV. The frequency region
junction to ensure no galvanic interaction between the of 0.01 Hz to 30 kHz was applied using a frequency response
dissimilar metals. The natural freshwater, the inhibitor treated analyzer. Each experiment was done with a replica. All the
freshwater and control water system were changed once in experiments were performed at ambient room temperature (30
every 24 h, up to 60-days in order to make the biofilm active. °C).
The measurement of peroxide in the biofilm was measured
using the peroxide strip (Merchoquant, Germany) method. 3. RESULTS AND DISCUSSION
EPS production was estimated by the standard procedure.14 3.1. Physico-chemical Analysis of Freshwater. The
2.3. Enumeration of Bacteria. The 60-day old natural physicochemical properties of freshwater are presented in Table
biofilm and the inhibitor treated biofilm on 316L SS were 2. Physico-chemical analysis of freshwater revealed that its pH
collected from the surface and washed with sterilized water to
remove the unattached microorganisms. The biofilm was Table 2. Physico-chemical Properties of Freshwater
scrapped using a sterilized knife and then 1 mL of biofilm
physico-chemical param. values
sample was directly mixed with 99 mL of sterilized water.
Various types of bacteria viz., heterotrophic bacteria (HB), acid temp. 30 °C
producing bacteria (APB), and manganese oxidizing bacteria turbidity 39.5 NTU
(MOB) were enumerated by pour plate method. The bacterial pH 7.4
density was expressed as colony-forming units per cm2 (CFU/ total dissolved solids 28.8 mg L−1
cm2) for all biofilm samples total suspended solids 49 mg L−1
2.4. Scanning Electron Microscopy. The biofilm covered dissolved oxygen content 6.9 mg L−1
specimen was washed with Milli-Q water to remove the conductivity 525 μS cm−1
unattached bacteria. The sample was fixed with 0.1 M chloride 79 mg L−1
phosphate buffer (pH 7.2) for 4 h at room temperature and total hardness 152 mg L−1
the specimens were immersed in 0.1 M phosphate buffer (pH sulfate 28 mg L−1
7.2) containing 2.5% gluteraldehyde + 4% paraformaldehyde manganese 0.9 mg L−1
copper 16 mg L−1
and kept overnight. Subsequently, the biofilm was dehydrated
iron 86 mg L−1
through a series of ethanol (25%, 50%, 75%, to 100%) and
acetone (100%) wash. The dehydrated biofilm attached
specimens were coated with gold sputter using ion sputtering was 7.4 with a dissolved oxygen concentration of 6.9 mg L−1.
(Jeol model JFC 1100) and biofilm surface was observed using The total dissolved solid content was 28.8 mg L−1. Chloride
SEM (Hitachi Model S-3000H). content was 79 mg L−1, whereas the sulfate content was 28 mg
2.5. FTIR Spectroscopy. Fourier-transform infrared L−1. The freshwater also contained 0.9 mg L−1 of manganese.
3.2. Enumeration of Bacteria. Table 3 shows the counts
(FTIR) spectra (Thermo Nicolet Nexus 670) were used for
of viable bacterial population in natural biofilm and biofilm
the analysis of the chemical characteristics of the biofilm
samples. The biofilm samples were ground into a fine powder
and pressed to pellet with the appropriate amount of KBr. The Table 3. Enumeration of Total Viable Bacterial Counts on
spectrum was taken in the mid-IR region of 400−4000 cm−1 the Natural Biofilm Covered over 316L SS (60 days old) and
and recorded in transmittance mode. ATMP + ZnSO4 Treated Biofilm Covered on 316L SS (60
2.6. Electrochemical Measurements. Open-circuit po- days old) at 30 °C
tentials (OCP) for 316L SS specimens in freshwater, inhibitor bacterial counts (CFU/cm2)
treated water, and sterilized freshwater (control) were system HB MOB APB
measured with time using a digital multimeter (Rish 18S
natural biofilm 4.6 × 106 3.7 × 106 3.0 × 105
Multimeter, India) against saturated calomel electrode (SCE)
ATMP + ZnSO4 treated biofilm 3.2 × 102 3.1 × 101 3.6 × 103
as reference electrode. Polarization experiments were con-
ducted in the respective systems under air-saturated condition
using a computer-controlled potentiostat (DC105, Gamry
instruments Inc., U.S.A.) in a one-liter corrosion cell. A formed in the presence of inhibitor (industrial biofilm) on 316L
three-electrode setup was used, which consisted of a 316L SS SS. The ranges of bacterial count measured were between 3.0 ×
specimen as a working electrode, SCE as the reference 105 and 4.6 × 106 CFU/cm2 in natural biofilm at 30 °C whereas
electrode and platinum foil as the auxiliary electrode. Cathodic in inhibitor treated biofilm, the bacterial count ranged between
and anodic polarization experiments were separately carried out 3.1 × 101 and 3.6 × 103 CFU/cm2 at 30 °C. It indicates that the
from OCP of the 316L SS and polarized to −1.0 V and +1.2 V bacterial density in the industrial biofilm was lower when
(SCE) at a scan rate of 0.166 mV s−1, respectively. The cyclic compared to natural biofilm. ATMP inhibitor plausibly controls
voltammetry (CV) experiments were carried out in the range the rate of bacterial growth in inhibitor treated biofilm covered
from −0.5 to 1.0 V (SCE) at a scan rate of 5 mV s−1. The on the surface of 316L SS specimen. The results show that
alternating current (AC) impedance studies were carried out ATMP has a fairly antibacterial effect.15
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Industrial & Engineering Chemistry Research Article

3.3. Scanning Electron Microscopy. Microscopic ob-

servation indicated that the natural biofilm contained different
microbial populations, including rod-shaped bacteria and
diatoms. SEM images also showed that the extent of
colonization with a higher number of organisms found in 60-
day old natural biofilm, often forming clumps of cells
heterogeneously distributed on the 316L SS surface (Figure
1a). A patchlike accumulation of biofilm matrix on stainless

Figure 2. FTIR spectrum of biofilm formed on 316L SS (60 days old)

(a) natural biofilm and (b) inhibitor (ATMP + ZnSO4) treated
biofilm.

the band at 1085 cm−1 corresponding to the presence of

polysaccharides and the other bands indicating amide I at 1650
cm−1 and amide II at 1550 cm−1 was also observed. Since the
band at 1450 cm−1 corresponds to amide I and amide II, it is
more likely that this band indicates methyl group (CH3) in
proteins. There was no inorganic band region observed in the
inhibitor treated system, which confirms the absence of
inorganic cations, nevertheless, trace organic attachment was
observed.
3.5. OCP Measurements. Figure 3 shows the OCP versus
exposure time of 316L SS by an average of five samples in each

Figure 1. SEM images of biofilm formed on 316L SS (60 days old) (a)
natural biofilm and (b) inhibitor (ATMP + ZnSO4) treated biofilm.

steel surface was observed. It is clearly observed that some

bacteria adsorb on 316L SS in the inhibitor treated biofilm. In
inhibitor treated biofilm, destined well-defined individual cells
can be noticed and (Figure 1b) there was no trace of a matrix in
the biofilm on 316L SS surface.
3.4. Fourier Transform Infrared Spectroscopy. Figure 2
shows the FTIR spectra of natural biofilm and inhibitor
(ATMP + Zn2+) treated biofilm. FTIR spectra (Figure 2a) of
natural biofilm shows peaks present in the range 429−795 cm−1
indicates MnOx stretching, bending, and wagging vibrations.16
A prominent band at 1035 cm−1 indicates the presence of
polysaccharides in natural biofilm. A peak around 1383 cm−1
indicates CH2 vibrations of polysaccharides. A band at 1640
cm−1 was recorded and indicates the polysaccharide, corre-
sponding to amide being indicative in the biofilm. The peaks at
around 1640 cm−1 and 3406 cm−1 were the OH-stretching
vibration and bending, respectively. FTIR analysis revealed that
the MnOx, OH stretching, and EPS fraction dominate the bulk
of natural biofilm. Hence, it can be concluded that the
electrochemical behavior of natural biofilm is due to the
combination of MnOx and organic molecules of biofilm. An
FTIR spectrum (Figure 2b) of inhibitor-treated biofilm shows
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Figure 3. Open-circuit potential (OCP) vs time for 316L SS with (a)
natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated biofilm, and
(c) control.
of the systems viz., natural biofilm, inhibitor (ATMP + Zn2+)
treated biofilm and control. The initial potential was about
−130 mV and the natural biofilm (Figure 3a) on 316L SS
gradually shifted toward positive direction about +380 mV.
While on adding inhibitors ATMP and Zn2+, the initial
potential was −30 mV but the positive shift could not be
noticed significantly (Figure 3b). In the control system, a slight
shift to a positive range was noticed from −160 mV to −50 mV.
It reveals that natural biofilm encourages the ennoblement
process, which supports the observation made by previous
dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181
Industrial & Engineering Chemistry Research
investigators,4,5,7,8,11,17,18 whereas the inhibitor treated (indus-
trial) biofilm did not encourage the ennoblement process.
Johnsen and Bardal19 and Lin and Dexter20 related this positive
process to the increased current density requirement for
cathodically protecting stainless alloys. This process was
correlated with pitting probability of stainless alloys, but
nobody has correlated this ennoblement process with industrial
biofilm that is inhibitor added system. This ennoblement
process was correlated with depositing of manganese by
manganese oxidizing bacteria on SS.7,21 It can be concluded
that since ATMP acts like a chelator, it may reduce the positive
shift of 316L SS.
3.6. Polarization Studies. 3.6.1. Cathodic Polarization.
The cathodic polarization curves for 316L SS covered with
natural biofilm, inhibitor treated biofilm covered on 316L SS,
and control are shown in Figure 4. The cathodic polarization
Figure 4. Cathodic polarization of 60-days old biofilm formed on 316L
SS (a) natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated biofilm,
and (c) control.
curve of 316L SS covered with natural biofilm exhibits two
reduction peaks (Figure 4a) which are similar to the peaks
noticed by electrochemically deposited manganese oxides on
the graphite electrode11 and chemically coated manganese
oxides.7 The result reveals that the presence of manganese in
natural biofilm encourages the reduction peak, whereas the high
oxygen reduction current, is due to the interaction between
H2O2 and manganese in the biofilm.7,8,11,18 The change of open
circuit potential in the electropositive direction can be
explained by an increase in the rate of the cathodic reaction.22
In the control system, no peak has been noticed and the
limiting current was lower when compared to 316L SS covered
with natural biofilm. The inhibitor treated system does not
contain any peak but the limiting current is lower (Figure 4b),
when compared to other systems. The reduction of cathodic
current in inhibitor added system is due to the adsorption of
ATMP and Zn2+. Since that inhibitor (ATMP + Zn2+) is added
in cooling water system, the mixed inhibitor component
reduces the cathodic current but does not encourage the
ennoblement process. It can also be claimed that Zn2+ may
adsorb on the surface of 316L SS, which reduces the cathodic
current. It can also be inferred that Zn(OH)2 formed on the
metal surface retards the oxygen reduction reaction and thus
controls the cathodic reaction of 316L SS.23 The interaction
between biogenic H2O2 and manganese oxide enhances the
limiting current by the redox behavior of the natural biofilm.11
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3.6.2. Anodic Polarization. Figure 5 shows the anodic
polarization curves for 316L SS covered with natural biofilm,
Figure 5. Anodic polarization of 60-day old biofilm formed on 316L
SS (a) natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated biofilm,
and (c) control.
inhibitor incorporated biofilm, and control. In the control
system, the breakdown potential for 316L SS was 1.0 V vs SCE.
The breakdown potential for natural biofilm and inhibitor
treated biofilm was most probably the same where as the
current for breakdown potential was lower when compared to
the control. The ip was higher in natural biofilm covered 316L
SS when compared to the control and inhibitor treated system.
This is due to the shifting of potential to the positive side by
natural biofilm. In the presence of an inhibitor treated system,
the polarization curve (Figure 5b) shifted toward the region of
lower current density (ip) among the polarization curves. It
indicates that inhibitor reduces ip of 316L SS when compared
to the other systems. It is due to the role of anodic inhibitor on
the metal surface of 316L SS. The reduction of anodic current
at all the potentials in natural biofilm and inhibitor treated
biofilm are also lower when compared to the control (without
biofilm). It can be concluded that biofilm and inhibitor treated
system on 316L SS reduces the corrosion process by adsorption
of anodic species.
3.7. Cyclic Voltammograms. Cyclic voltammograms of
the natural biofilm, inhibitor treated biofilm, and control for the
316L SS are presented in Figure 6. Three peaks were noticed in
the natural biofilm (Figure 6a), two peaks at the anodic region
(oxidation) and another peak at cathodic region (reduction).
Long et al.24 noticed the oxidation and reduction peak of nano
mesoporous MnOx in aqueous electrolyte at +240 mV and −15
mV (Ag/AgCl) respectively, which are similar to the present
observation. The redox behavior of biogenic MnOx is due to
the formation of an oxidation peak (I) and a reduction peak
(III) at +248 mV and 0 mV (SCE), respectively.11 The
difference in potential of the redox peaks (ΔEp) was
approximately +248 mV as observed from the CV response
of natural biofilm. It was confirmed that MnOx redox couple
was a quasi-reversible system. Another peak (II) in the anodic
side (+700 mV) was noticed and identified as H2O2 peak. This
observation supports with the observation made by Yao et al.25
who noticed peroxide a peak at +700 mV (SCE) by using GCE
in phosphate buffer at pH 7.4. The peroxide was also estimated
by the appearance of blue color on peroxide strip and the
concentration of peroxide was in the range 3−5 mg L−1 with
exposure time.
dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181
Industrial & Engineering Chemistry Research
Figure 6. Cyclic voltammograms of 60-day old biofilm formed on
316L SS (a) natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated
biofilm, and (c) control.
Whereas inhibitor (ATMP + Zn2+) treated biofilm covered
316L SS was not observed with redox peaks, the anodic current
was slightly increased at +700 mV (SCE). The peroxide was
also observed in inhibitor treated biofilm (3−5 mg L−1) by
peroxide strip method. The CV of the inhibitor treated biofilm
reveals that there is no accumulation of manganese oxide on
inhibitor treated biofilm covered 316L SS (Figure 6b). Since
ATMP inhibitor removes all the cations in the biofilm, the
redox peaks in cyclic voltammogram could not be noticed.
3.8. AC Impedance Analysis. Figures 7, 8, and 9 show the
Nyquist plot, the Bode plot, and the proposed equivalent
Figure 7. Nyquist plots of 60-day old biofilm formed on 316L SS (a)
natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated biofilm, and
(c) control.
circuit, respectively, for natural biofilm and inhibitor treated
(industrial) biofilm covered 316L SS and control, together with
their numerical fittings. The equivalent circuit consisted of
parameters characterizing the electrolyte resistance (Rs), charge
transfer resistance (Rct), capacitance (Cdl) film resistance (Rf),
and Warburg diffusion impedance (ZW). The parameters fitted
using Gamry software version 3.2 are listed in Table 4. The Rs
value for the 316L SS exposed to natural freshwater and
inhibitor treated freshwater was higher when compared to the
control system. While adding inhibitor, Rs value was higher
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Figure 8. Bode plots of 60-day old biofilm formed on 316L SS (a)
natural biofilm, (b) inhibitor (ATMP + ZnSO4) treated biofilm, and
(c) control.
Figure 9. Equivalent circuit model of 316L SS with (a) natural biofilm,
(b) inhibitor (ATMP + ZnSO4) treated biofilm, and control.
when compared to the natural biofilm. The attachment of
microorganisms and adsorption of inhibitors with biofilm
enhance the Rs value. Table 3 shows the high count of bacteria
in natural biofilm when compared to inhibitor treated biofilm.
The natural biofilm consists of more mucous substance
(matrix−organometallic complex); separated and well-defined
cells can be noticed in the inhibitor treated biofilm. In the
present study, the Cdl value of the control system was 114 μF
cm−2, whereas in natural biofilm, the Cdl value increased to
1680 μF cm−2. Inhibitor treated biofilm had 39.1 μF cm−2. This
result suggests that Cdl is related to the Helmholtz double layer
capacitance associated with the passive film/solution interface.
The decrease in electrical capacitance with an increase in oxide
film thickness might possibly lie in the properties of the
aqueous environment. The increase in Cdl value in natural
biofilm is due to the availability of more free charges in the
biofilm and reduces the oxide film thickness. The heteroge-
neous nature of biofilm also determines Cdl value of 316L SS.
The adsorption of inhibitor on 316L SS increases the thickness
of the film and reduces the Cdl value.
The equivalent circuit of 316L SS covered with natural
biofilm (Figure 9a) shows the passive film represented by the
constant phase element (CPE), ZCPE in parallel with film
resistance (Rf). The resistance of the passive film on the 316L
SS decreased when exposed in natural freshwater biofilm. The
resistance of oxide film on 316L SS in control system was 2.39
× 166 Ω cm2, which reduced to 1.16 × 104 Ω cm2 in the
presence of natural biofilm. It is due to the disturbance of oxide
film by the natural biofilm which enhances corrosion.26 While
adding inhibitor treated system, the film resistance of 316L SS
was 2.06 × 1014 Ω cm2. It is due to the adsorption of inhibitor
over the oxide film of stainless steel. The inhibitor treated
system has higher film resistance27 when compared to natural
biofilm, which is due to the adhesion of the inhibitor monolayer
dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181
Industrial & Engineering Chemistry Research
Table 4. Impedance Parameters of Equivalent Circuits Representing, 316L SS Exposed in Freshwater (Natural Biofilm),
Inhibitor (ATMP + ZnSO4) Treated Biofilm, and Control at 30 °C for a Duration of 60 days
system OCP (mV vs SCE) Rs (Ω cm2) Cdl (F cm−2)
−4
control −50 mV 313 1.14 × 10
natural biofilm +380 mV 655 1.68 × 10−3
inhibitor treated biofilm +20 mV 1179 3.91 × 10−5
on the 316L SS surface. Warburg impedance (ZW) in the
equivalent circuit was introduced to account for the diffusion
process within the system (Figure 9a). Bode plot, the phase
angle shows 45° shift, indicating the diffusion process in the
system. The diffusion process is due to the interaction between
biogenic manganese and peroxide in the natural biofilm, which
determines the diffusion process. This concept is supported by
the cathodic polarization curve where the reduced species of
manganese and ferric in natural biofilm determine the diffusion
mechanism. A tiny semicircle (Figure 7a) in Nyquist plot
appearing at the high frequencies11,28 may be related to an
electrochemical reaction involving biogenic MnOx (eq 1).
MnO2 + H+ + e− ⇌ MnOOH (1)
The interaction between biogenic MnOx and peroxide does
not influence the diffusion process in inhibitor treated
(industrial) biofilm. This observation was made by low
frequency region of the Nyquist plot (Figure 7b). ATMP and
Zn2+ inhibitor film formation of the oxide film was represented
by CPE and Rf in the equivalent circuit (Figure 9b).
4. CONCLUSIONS
The present study explains the electrochemical behavior of
natural biofilm and inhibitor treated biofilm on 316L SS. The
inhibitor added biofilm covered on the 316L SS reduced
cathodic current density when compared to the natural biofilm
and control. Anodic polarization revealed that ip is higher in
natural biofilm when compared to inhibitor treated biofilm. The
CV result of natural biofilm covered 316L SS revealed the redox
behavior of biogenic MnOx. Since ATMP inhibitor removes all
cations in the biofilm, the redox peaks in cyclic voltammogram
could not be noticed in inhibitor treated (industrial) biofilm.
Electrochemical impedance spectroscopy revealed that a
protective film was formed on the 316L SS surface when
exposed to an inhibitor treated freshwater. It can be claimed
that the inhibitor treated biofilm does not encourage the pitting
probability and also reduces the corrosion current. This study
also claims that inhibitor added biofilm does not encourage
ennoblement phenomenon in cooling water system.
■ AUTHOR INFORMATION
Corresponding Author
*Tel.: +91-4565-241400. E-mail: swamy24@gmail.com.
Notes
The authors declare no competing financial interest.
■ ACKNOWLEDGMENTS
The authors are grateful to the Director, CSIR-CECRI,
Karaikudi for his encouragement and facilities provided.
■
REFERENCES
(1) Pedersen, A.; Hermansson, M. Inhibition of metal corrosion by
bacteria. Biofouling 1991, 3, 1−11.
16180
Article
ZCPE
Rct (Ω cm2) Yo (Sn/ Ω) n1 Rf (Ω cm2) W (Ω−1)
−4
967 3.67 × 10 0.80 2.39 × 10 6
2348 1.21 × 10−3 0.65 1.16 × 104 2.39 × 10−5
3377 1.23 × 10−4 0.81 2.06 × 1014
(2) Maruthamuthu, S.; Rajagopal, G.; Sathiyanarayanan, S.;
Angappan, S.; Eashwar, M.; Balakrishnan, K. Contributions of oxide
film and bacterial metabolism to the ennoblement process: Evidence
for a novel mechanism. Curr. Sci. 1996, 71, 315−320.
(3) Jeyaraman, A.; Earthman, J. C.; Wood, T. K. Corrosion ion by
aerobic biofilms on SAE 1018 steel. Appl. Microbiol. Biotechnol. 1997,
47, 62−68.
(4) Mollica, A.; Trevis, A. France Proceeding 4th International Congress
on Marine Corrosion and Fouling. Juans-les-Pins, Antibes, France, 1976;
351−365
(5) Eashwar, M.; Maruthamuthu, S.; Sathiyanarayanan, S.;
Balakrishnan, K. The ennoblement of stainless alloys by marine
biofilms: The neutral pH and passivity enhancement model. Corros.
Sci. 1995, 37, 1169−1176.
(6) Olesen, B. H.; Avci, R.; Lewandowski, Z. Manganese dioxide as a
potential cathodic reactant in corrosion of stainless steels. Corros. Sci.
2000, 42, 211−227.
(7) Dexter, S. C.; Maruthamuthu, S. Response of passive alloys with
n- and p-type passive films to manganese in biofilms. Corrosion; NACE
International: Houston, TX, 2001; No. 01256.
(8) Lewandowski, Z.; Hamilton, A. MIC of stainless steels as a model
system to study metal-microbe interactions. Corrosion; NACE
International: Houston, TX, 2002; No 02474.
(9) Ford, T. E.; Walch., M.; Mitchell, R. Corrosion of metals by
thermophilic microorganisms. Mater. Perform. 1987, 26 (No. 2), 35−
39.
(10) Mohanan, S.; Maruthamuthu, S.; Kalaiselvi, N.; Palaniappan, R.;
Venkatachari, G.; Palaniswamy, N.; Raghavan, M. Role of quaternary
ammonium compounds and ATMP on biocidal effect and corrosion
inhibition of mild steel and copper. Corrosion Rev. 2011, 23, 425−444.
(11) Sridharan, D.; Manoharan, S. P.; Palaniswamy, N. Redox
behavior of biofilm on glassy carbon electrode. Bioelectrochemistry
2011, 82, 135−139.
(12) Clesceri, L. S.; Greenberg, A. E.; Rhodes Trusselil, R. Standard
Methods for the Examination of Water and Wastewater, 17th ed.;
American Public Health Association (APHA) and American water
works (AWW): Washington, DC, 1989.
(13) Trivedy, R. K.; Goel, P. K. Chemical and Biological Methods for
the Water Pollution Studies; Environmental Publications: Karad, India,
1986.
(14) Dubois, M.; Gilles, K. A.; Hamilton, J. K.; Rebers, P. A.; Smith,
F. Colorimetric method for determination of sugars and related
substances. Anal. Chem. 1956, 28, 350.
(15) Najoua, L.; Mounim, L.; Fouad, B.; Nour-Eddine, C.; Souad El,
H.; Charafeddine, J. Corrosion inhibition of carbon steel and
antibacterial properties of aminotris-(methylenephosphonic) acid.
Mater. Chem. Phys. 2010, 119, 330−336.
(16) Parikh, S. J.; Chorover, J. FTIR spectroscopic study of biogenic
Mn-oxide formation by Pseudomonas putida GB-1. Geomicrob. J. 2005,
22, 207−218.
(17) Maruthamuthu, S.; Sathiyanarayanan, S.; Palaniappan, R.;
Palaniswamy, N. Role of manganese depositors on ennoblement
process: Evidences for adsorption theory. Corrosion; NACE Interna-
tional: Houston, TX, 2002; Paper No. 02468.
(18) Chandrasekaran, P.; Dexter, S. C. Mechanism of potential
ennoblement on passive metals by seawater biofilms. Corrosion; NACE
International: Houston, TX, 1993; No. 493.
(19) Johnsen, R.; Bardal, E. Cathodic properties of different stainless
steel in natural seawater. Corrosion 1985, 41 (5), 296−302.
dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181
Industrial & Engineering Chemistry Research Article

(20) Lin, S. H.; Dexter, S. C. Effects of temperature and magnesium

ions on calcareous deposition. Corrosion 1988, 44 (No. 9), 615.
(21) Dickinson, W. B.; Caccavo, F.; Lewandoski, Z. The
ennoblement of stainless steel by manganic oxide biofouling. Corros.
Sci. 1996, 38, 1407−1422.
(22) Rhoads, A.; Beyenal, H.; Lewandowski, Z. A microbial fuel cell
using anaerobi respiration as ananodic reaction and biomineralized
manganese as a cathodic reactant. Environ. Sci. Technol. 2005, 39,
4666−4671.
(23) Gopi, G.; Manimozhi, S.; Govidaraju, K. M.; Manisankar, P.;
Rajeswari, S. Surface and electrochemical characterization of pitting
corrosion behavior of 304 stainless steel in ground water media. J.
Appl. Electrochem. 2007, 37, 439−449.
(24) Long, J. W.; Young, A. L.; Rolison, D. R. Spectroelectrochemical
characterization of nanostructured, mesoporous manganese oxide in
aqueous electrolytes. J. Electrochem. Soc. 2003, 150, A1161−A1165.
(25) Yao, S.; Yuan, S.; Xu, J.; Wang, Y.; Luo, J.; Hu, S. A hydrogen
peroxide sensor based on colloidal MnO2/Na-montmorillonite. Appl.
Clay Sci. 2006, 33, 35−42.
(26) Hiromoto, S.; Hanaw, T. Electrochemical properties of 316L
stainless steel with culturing L929 fibroblasts. J. R. Soc. Interface 2006,
3, 495−505.
(27) Rajendran, S.; Apparao, B. V.; Palaniswamy, N.; Amalrajv, A. J.;
Sundarvadivelu, M. The role of phosphonates as transporters of Zn
ions in the inhibition of carbon steel in neutral solutions containing
chlorides. Anticorros. Method Mater. 2002, 49, 205−209.
(28) Ding, K. Q. Cyclic voltammetrically-prepared MnO2 coated on
an ITO glass substrate. J. Chin. Chem. Soc. 2009, 56, 175−181.

16181 dx.doi.org/10.1021/ie401279m | Ind. Eng. Chem. Res. 2013, 52, 16175−16181