Electrochimica Acta 54 (2008) 74–79

Contents lists available at ScienceDirect

Electrochimica Acta
journal homepage: www.elsevier.com/locate/electacta

Biocorrosion of stainless steel grade 304L (SS304L) in sugar cane juice

S. Durmoo a,b , C. Richard a,∗ , G. Beranger a , Y. Moutia b
a
Université de Technologie de Compiègne (UTC), Laboratoire Roberval, UMR CNRS 6253, BP 20529, 60205 Compiègne cedex, France
b
Mauritius Sugar Industry Research Institute (MSIRI) and University of Mauritius (UoM), Le Reduit, Mauritius

a r t i c l e i n f o a b s t r a c t

Article history: The main objective of this research study is to assess the behaviour of stainless steel AISI 304L in contact
Received 7 November 2007 with sugar cane juice. Stainless steel grade 304L is largely used in the sugar cane industry but the acidic
Received in revised form 6 May 2008 nature of sugar cane juice pose a serious challenge in maintaining the life span of vital components.
Accepted 10 June 2008
Sugar cane juice is acid having a pH value of around 5.6 at extraction. This acidic property is accounted
Available online 24 June 2008
for by the presence of a variety of acids namely: aconitic, citric, malic, oxalic, glycolic, mesaonic, tartic,
succinic, fumaric and syringic in sugar cane juice.
Keywords:
In addition to these acids there are approximately 50 different kinds of microorganisms present in the
Cane sugar juice
Austenitic stainless steel
green cane and which are very active. These microorganisms will act as a contributor to a rather quick
Pitting drop in pH (pH∼3.1) of the sugar cane juice once extracted.
Biocorrosion The more so, several minerals like water, salt, sulphate and silica are also present throughout the process
Biofilm line and are other contributor in the surface degradation wear mechanism.
Faced with all these adverse elements, it is therefore fundamental to investigate thoroughly in the wear
corrosion mechanism and biocorrosion on stainless steel grade 304L.
To evaluate the mass loss, several corrosion experimentations were carried with the help of a poten-
tiostat both in a sterilized juice and none sterilized juice. From these experimentations, it has been noted
that corrosion was present on the surface of the disc (SS304L) in contact with none sterilized juice in the
form of pitting while no corrosion wear was observed in the case of sterilized juice.
The presence of biofilm was also observed on the sample disc surface. Biofilm formed on the surface of
the sample disc was transferred to a potato dextrose agar (PDA) plate by stamping the disc in a circular
sequence on the plate in sterilized condition in view to measure the density and resistance of the film.
It was noted through this stamping protocol that the biofilm was very resistant due to the fact that after
12 stamping time we still observe the trace of the biofilm on the PDA plate. Microorganisms present in
the biofilm have growth on the PDA plate and isolation of each colony was carried out in view of their
identification.
© 2008 Elsevier Ltd. All rights reserved.

1. Introduction cane juice tends to favour corrosion to many types of equipment.

Stainless steels of austenitic type are well known in its capacity
to preserve the structure at ordinary temperature and are usu-
ally called 18-8 or 18-10 according to the percentage of chromium
and nickel. These stainless steels grades are heavily used in the
sugar industry and agro industry. Their relative facility in fabrica-
tion and their aptitude of deformation and also their good corrosion
resistance are so many reasons of using them in various fields,
particularly in the sugar industry [1].
Indeed, the sugar industry installations can be particularly sub-
ject to corrosion phenomena. At every step along the process line
(from crushing to crystallisation of sugar) the composition of the
∗ Corresponding author. Tel.: +33 3 44 23 46 17; fax: +33 3 44 23 49 84.
The types of corrosion recorded are of wide range (with no prefer-
ence order): bacterial corrosion, stress corrosion, corrosion under
deposit, intergranular corrosion, fatigue corrosion and pitting cor-
rosion. All of these corrosion phenomena are also present in the
beet sugar industry [2].
The first work mentioning corrosion phenomenon in the sugar
industry (beet sugar industry) in France goes back to the years
1950, more particularly at the stages process of the diffusion
and evaporation, critical step in the manufacture of sugar (beet
sugar).
Among the most widespread forms of corrosion, the localised
pitting corrosion is certainly the most frequent.
Sugar cane juice is a complex medium due to the continuous
fermentation process occurring once extracted and as well as the
presence of others mechanisms in the juice: such as chemical,

0013-4686/$ – see front matter © 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.electacta.2008.06.028
 S. Durmoo et al. / Electrochimica Acta 54 (2008) 74–79 75

Fig. 1. Schematic diagram of the jar device used to verify the growth of biofilm on
our specimen at the laboratory of MSIRI.
Fig. 2. Photo showing the specimen in the flask, before and after pouring fresh juice.

microorganism (bacteria, yeast and fungi), abrasion due to the

presence of suspended solids along the process line, etc. getting in contact (i.e. flask and specimen) with fresh sugar cane
To assess each of these mechanisms on the behaviour of stainless juice.
steel grade 304L, different kind of measurements and techniques The flasks were then placed on an arbitral shaker (Gallenghamp
were required: electrochemical measurements for the understand- model) set at 100 rpm for 2 days at a constant temperature of 25 ◦ C
ing of the corrosion phenomena, the transfer of biofilm formed on to allow a continuous mixing of the juice in view to keep the solu-
the samples to identify bacterias and yeast, the use of tribocorro- tion homogeneous (Fig. 3).
sion device to simulate erosion corrosion and abrasion corrosion After a period of incubation of 2 days, the specimen was removed
phenomenon. from the flask under the laminar flow cabinet to prevent exter-
Throughout this research work, substantial amount of results nal contamination. It was then rinsed in sterile distilled water and
have been compiled on the behaviour of SS304L surface in contact treated in an ultrasonic bath (Branson model) for 5 min. This was
with the sugar cane juice for the different degradation mechanisms done in order to ensure that only a strongly adhered biofilm would
(uniform corrosion, localised pitting corrosion, biocorrosion, and remain on the specimen.
erosion–abrasion corrosion). However, in this article only the bio- The technique of transferring the biofilm from the specimen to
corrosion and more particularly pitting results are presented. Some the culture medium was carried out according to a predefined pro-
results for uniform corrosion are also given. tocol which consisted of imprinting the specimen on the PDA plate
in a clockwise direction under sterilize atmosphere as illustrated in
Fig. 4. This imprinting procedure was repeated four times on a PDA
2. Methodology
Petri dish. In order to estimate the extent and tenacity with which
the biofilm was adhering to the specimen, a total of 12 prints were
In this research work, while studying the corrosion behaviour
made with a single specimen on a total of three PDA Petri dishes.
of specimens (SS304L) in contact with the sugar cane juice, the
To evaluate the impact of corrosion rate, electrochemical tests
presence of biofilm was observed on the exposed surface and more
were performed on fresh, aged and sterilized sugar cane juice
particularly in the region where pitting corrosion was occurred. It
jointly at the University of Mauritius and at the laboratory Rober-
was therefore essential to transfer the biofilm to a medium that
val (UTC). The juice was obtained from the MSIRI by high-pressure
promoted the growth of these microorganisms in order to iden-
extraction (2 bars) of cane chips using a French press model.
tify (and observe) their morphology and aid in their identification.
Throughout the duration of these tests, corrosion potential was
The potato dextrose agar (PDA) medium was chosen as it is a rich
measured against time. The polarisation curves were also recorded
medium in terms of nutrient composition and has the potential of
in the range of ±1500 mV/SSE for a sweep rate of 2.5 mV s−1 . The
promoting the rapid growth of microorganisms. It is nonselective
and will therefore allow a large array of microorganisms to grow.
This investigation of the biofilm and the identification of microor-
ganisms were carried out at the Mauritius Sugar Industry Research
Institute (MSIRI).
Fresh and sterilized sugar cane juice was poured separately in
500 ml sterile flask and sealed with a cotton plug. The specimen
(SS304L) of polished surface of grain 800 ␮m having a diameter
of 10 mm was first cleaned with 70% alcohol followed by several
washes in distilled water prior to sterilization at 119 ◦ C for 35 min
in order to eliminate the presence of foreign matter, bacteria and
other microorganisms on the surface. It was then placed in a vertical
plane and immersed in the juice solution at a specific depth (15 cm)
so as to standardise experimental conditions across every experi-
ment. The purpose of placing the specimen in a vertical plane in the
solution was to ascertain that adhesion of microorganisms was due
to their ability to form a biofilm against the natural downward force
of attraction due to gravity. Fig. 1 illustrates the principle described Fig. 3. Fresh juice in sterilised jar and placed on an arbitral shaker at constant
above and the photo in Fig. 2 shows the specimen before and after temperature of 25 ◦ C.
76 S. Durmoo et al. / Electrochimica Acta 54 (2008) 74–79

Fig. 4. Schematic diagram showing the technique of transferring biofilm from spec- Fig. 5. Evolution of the extracted sugar cane juice pH (cane category M695/69) with
imen of SS304L to the PDA medium in sterilised condition. time at ambient temperature.

Table 1
Definition of electrochemical parameters used to study corrosion, erosion corrosion
and abrasion corrosion of stainless steel 304L in contact with sugar cane juice

Item Description Parameters

WE Working electrode Area 0.73 cm2 for

corrosion and 0.58 cm2
for erosion
corrosion/abrasion
corrosion
RE Reference electrode Hg/Hg2 SO4
(SSE = +408 mV/SCE at
25 ◦ C)
AE Auxiliary electrode Platinum sheet
Medium Sugar cane juice pH range 5.6–3.4 for
corrosion and pH 5 for
erosion
corrosion/abrasion
corrosion
E Applied potential ±1500 mV Fig. 6. Semi-logarithmic scale representing overlapped polarization curves (−1500
T Sweep rate 2.5 mV/s to +1000 mV) with a sweep rate 2.5 mV/s for some pH values of sugar cane juice
solution (fresh and aged) and a control sterilised juice at ambient temperature.

reference electrode (RE) used for all electrochemical tests (study
of corrosion, erosion corrosion and abrasion corrosion) was an
Hg/Hg2 SO4 reference electrode (SSE) intended for the applications
where chloride could interfere with the measurement. It is noted
that the reference potential at 25 ◦ C for an electrode SSE filled with
saturated K2 SO4 solution is approximately +408 mV with respect to
a saturated calomel electrode (SSE = +408 mV/SCE). The test speci-
men, of cylindrical shape, was used as the working electrode WE.
A platinum sheet was used as our auxiliary electrode AE. All these
data are summarised in Table 1. Potentiostat used in this research
work was a Volta Lab PGP 201.
It is also important to have in mind that the acid content of
sugar cane juice depends on its geological place and its variety. In
this study, the commercial variety M695/69 was used for the whole
study and the extracted juice was supplied by the MSIRI.
3. Results and discussion
Knowing that the sugar cane juice was a complex and an evolu-
tionary medium, it was essential to characterize its variation with
time by measuring its pH evolution. Fig. 5 shows a typical pH evo-
lution of the sugar cane juice extracted from variety M695/69 with
time. Fresh sugar cane juice has a pH value of around 5.4; this value
and this curve may vary depending on the variety of sugar cane, its
growth stage and also on the geographical location and environ-
mental conditions. Step by step, the pH value dropped to about 3.1.
Bacteria, yeast and fungi are among the three main elements in this
complex microorganism system, responsible for this drop in pH as
already stated in some published papers [3–6].
To assess the corrosion impact on the surface of SS304L, elec-
trochemical experimentation was carried out with sugar cane juice
of variety M 695/69 of different pH value at ambient temperature
and also with sterilized juice to identify the effect of bacteria. The
results are presented in Fig. 6. It is clearly demonstrated from the
evolution of the corrosion potential (Ecorrosion ) with pH that when
the juice pH is reduced the specimen SS304L becomes less resistant
against corrosion. With sterilized juice the pH value remains stable;
this is due to the fact that all microorganisms present in the solu-
tion have been eliminated and in terms of corrosion the specimen
SS304L exhibits very small surface degradation.

Table 2
Summary of thickness loss of results of SS304L in contact with sugar cane juice of pH ranging from 5.6 to 3.6 (corrosion scenario)

Case Type of test pH Ecorrosion (mV/SSE) Icorrosion (␮A/cm2 ) Thickness loss e (mm/year)

A Corrosion: fresh sugar cane juice 5 −445 8.38 0.098

B Corrosion: aged sugar cane juice 3.6 −575 12.40 0.145
C Corrosion: sterilized juice 5.6 −500 0.50 0.006
 S. Durmoo et al. / Electrochimica Acta 54 (2008) 74–79
Fig. 7. Observation (ESEM) pitting corrosion on SS304L surface in contact with fresh sugar cane juice of pH 5 under polarisation test (−1500 to +1000 mV) with a sweep rate
of 2.5 mV s−1 at ambient temperature.
Taking into account the evolution of the electrochemical curves
it can be emphasized that a modification of uniform corrosion can
occur. From curves of Fig. 6, the uniform corrosion rate in term of
thickness loss was estimated for corrosion scenario of stainless steel
SS304L in contact with sugar cane juice of pH ranging from 5.6 to 3.6
at ambient temperature (Table 2). The weight loss measurements
were not significant.
From Table 2, it is clearly shown that while pH drop the rate of
corrosion increases in agreement with different results obtained
in various media [3]. Let us note that insignificant metal loss is
observed in a sterilized medium (case C).
The observation of the metal surface (under environmental
scanning electron microscope (ESEM)) revealed the occurrence of
pitting phenomena on the sample surface while in contact with
the fresh juice of pH 5.6. Sugar cane juice contains a high level of
chloride (approximately 1300 ppm) [7,8]. Chlorides tend to become
concentrated with sugar processing [9]. Nevertheless, no pitting
was observed when same experimentation was performed with
sterilized juice pH 5.6. Fig. 7 shows pitting corrosion and presence of
biofilm around the pitting cavities after a polarisation test of −1500
to +1500 mV, at a sweep rate of 2.5 mV s−1 . Under same experimen-
Fig. 8. Absence of corrosion is noted on the surface of SS304L after a polarisation
test.
77
tal condition but with a sterilized medium (Fig. 8) no presence of
biofilm on the surface or pitting corrosion was observed. The pres-
ence of some crystal sugar which is due to the concentration of the
juice solution was only detected at a high magnification (8000×).
The fact that the presence of biofilm was observed on the sur-
face especially around the pitting perimeter and even in the pitting
hole then it is essential to investigate the presence of the biofilm
on the surface. Through the printing protocol as explained earlier
in Section 2, the biofilm was transferred from the SS304L speci-
men surface to the PDA plate and was maintained at 30 ◦ C in an
incubator. The experiments have been carried out with two sep-
arate samples: one in contact with fresh juice and the other with
sterilized juice both during a period of 1 day. There is clear dis-
tinct growth of microorganisms after 1 day of incubation period
as shown in Fig. 9(a) which confirms the presence of adhesion of
biofilm on the SS304L specimen surface in contact with fresh juice
medium.
Contrary to Fig. 9(a) there is no growth of microorganisms even
after 3 days of incubation period as shown in Fig. 9(b) where the
cane juice used was sterilised and therefore confirms the prediction
earlier formulated.
To verify the adhesion of the biofilm adhesion growing on the
surface of the specimen when fresh juice was used, the same pro-
cedure was repeated 12 times in total on three separate PDA plates
as shown in Fig. 10. It is clearly observed that even after 12 imprints
there is still residue of biofilm present on the surface of the sample.
Fig. 9. (a) Observation of microorganism growth on PDA medium for SS304L sample
in contact with fresh juice after an incubation period of 1 day at 30 ◦ C and (b) no
microorganism growth on PDA medium for SS304L in contact with sterilized juice
after an incubation period of 3 days at 30 ◦ C.
78 S. Durmoo et al. / Electrochimica Acta 54 (2008) 74–79

Fig. 10. Verification of the adhesion capacity of the biofilm on the sample SS304L: (a) the sample was imprinted four times on the first PDA medium under sterilized condition,
(b) four more imprinting was made from the same sample again under sterilised condition and (c) four other more imprinting was made with the same sample in sterilized
condition, and all three PDA plating was kept in an incubator for 1 day at 30 ◦ C.

From the imprints on PDA plates several types of colonies were

growing and the idea was to separate each colony without contam-
ination to obtain pure colonies and this technique is called isolation
of colonies. After repeating this exercise two to three times, a pure
colony is obtained as shown in Fig. 11. Once each colony is isolated,
the next step is to proceed to its identification and lyophilisation.
However, with experience yeast and bacteria can be easily distin-
guished by observing their shape and morphology with the aid of
a medium to high power lens of a light microscope.
During the phase of purification of colony (having well sepa-
rated colonies), secretion of pigmentation was observed in certain
cases on PDA plate after 3 days of incubation as shown in Fig. 12. At
least two types of colonies can be observed from this figure, secret-
ing each a specific acid content which can be distinguished by the
colour of the pigmentation on the plate.
Finally to put results beyond doubt concerning the adhesion of
the biofilm on the surface of SS304L, is to confirm visually and
Fig. 12. Secretion of pigmentation (red, blue and green colour) was observed on the
empirically the same specimen with the help of an ESEM. PDA plating after 3 days of incubation at 30 ◦ C. (For interpretation of the references
The same precautionary procedures as outlaid earlier in the to colour in this figure legend, the reader is referred to the web version of the article.)
transfer of the biofilm to the PDA plate were applied on the sam-
ple in contact with the sugar cane juice of variety M 695/69. In fact Fig. 14 which has been enlarged to higher magnification shows
the specimen was removed after 1 day of contact with the juice, the texture and arrangement of species growing on the surface of
washed with sterilized de-mineralised water and then observed the SS304L. It reveals in fact that the surface density of bacteria
under ESEM. covering a specimen surface is very high.
Fig. 13 under a magnification factor of 1000 times; reveal the A single bacterium observed at higher magnification (Fig. 15)
presence of microorganism adhering to the SS304L surface. reveals the presence of flagella which enable the bacterium to move
Different clustering colonies grouped together can be observed from one place to another within on the biofilm; this possibility of
and among which it is easy to distinguish yeast species which migration and consequently the extension of the biofilm across the
have the particular characteristics of their bright and round sur- specimen surface can explain the coating of the surface observed
faces. in Fig. 14.

Fig. 11. Purification process in view to separate each colony present in the biofilm without contamination: (a) a particular colony was collected from the imprinting PDA (a)
under sterilized condition and spread on this PDA plating and kept growth in incubator for 1 day at 30 ◦ C, (b) from plating (a) a small zone was collected under sterilised
condition and spread again on a new PDA plate and kept growth in incubator for another day at 30 ◦ C and (c) same procedure in (b) was repeated again and kept for 1 day in
an incubator at 30 ◦ C.
 S. Durmoo et al. / Electrochimica Acta 54 (2008) 74–79
Fig. 13. Bacteria and yeast colonies growing on the surface of SS304L observed under
ESEM.
Fig. 14. High surface density population growth of a colony on the surface of SS304L
observed under ESEM.
Fig. 15. Observation of a bacterium with its flagella on the surface of SS304L under
ESEM.
79
It is important to note that pitting observed is of the diameter
size of 20 ␮m in average, which can be considered as quite big. Then
the possibility of corollas corrosion phenomena as described in the
literature review [2] may have been produced here.
4. Conclusion
The findings of this study have shown that various forms of cor-
rosion were occurring when the SS304L specimens were in contact
with sugar cane juices as
(i) Uniform corrosion. Nevertheless, the presence of this kind of cor-
rosion was low and was depending on pH of the juices. The pH
dropped with time.
(ii) Pitting corrosion. Pitting phenomena generally occurred in fresh
or aged juice but not in sterilized juice. This corrosion grew
under the passive film due to a confinement and then gave
small craters on the surface of the specimens. These phenom-
ena arise from the complexity of the media with the presence of
chlorides and are also depending on time with a concomitant
decrease of pH. The question that can be raised: this pitting
corrosion is it chemical and electrochemical result or a bio-
chemical result or a coupling of the two mechanisms? Indeed,
bacteria can locally affect by their metabolism the passive
film.
Other types of corrosion were observed on surface of SS304L used
in sugar cane industry particularly as tribocorrosion phenomena
(abrasion–erosion corrosion). These results will be reported subse-
quently.
From a technological point of view, the factor time is an
important parameter and it is convenient to reduce the time of
contact with a given juice in order to prevent the effects of its
aging.
It is worth mentioning that the results presented are not exhaus-
tive but comprehensive enough to understand the behaviour and
mechanism of the biofilm in the degradation of stainless steel grade
304L.
Acknowledgements
This work was supported by the AUF (Agence Universitaire de la
Francophonie) under grant of Dr. Seeva Durmoo. The work was also
associated to the Biocorys programme and the authors are indebted
to Dr. Damien Féron from CEA Saclay (France), scientific supervisor
of the Biocorys programme for his very helpful advice.
References
[1] F. Dupoiron, M. Verneau, in: F. Dabosi, G. Béranger, B. Baroux (Eds.), Corrosion
Localisée, Ed. Sci., Les éditions de Physique, 1994, p. 605.
[2] C. Poiré, Phénomènes de corrosion dans l‘industrie sucrière: comportement de
l‘acier inoxydable à 17% de chrome dans les évaporateurs, thèse UTC, 1986.
[3] M.A. Clarke, Cane Hand Book, Sugar Processing Research INC., New Orleans
Louisiana, p. 18.
[4] W.L. Owen, Facts About Sugar 18 (1925) 566.
[5] W.L. Owen, Microbiology of Sugar Manufacture and Refining, p. 385.
[6] P.C. Irvine, Sugar Bull. 42 (23) (1964) 317.
[7] B. Baroux, C. Lemaître, F. Dabosi, in: P. Lacombe, B. Baroux, G. Béranger (Eds.),
Les aciers inoxydables, Ed. Sci., Les Editions de Physique, 1990.
[8] G.A. Cash, D.P. Schweinsberg, G.A. Hope, Corros. Sci. 30 (6, 7) (1990) 543.
[9] G.A. Cash, G.A. Hope, D.P. Schweinsberg, Corros. Sci. 33 (5) (1992) 751.