Polish Journal of Chemical Technology,

Pol. J. Chem.
20, 3,
Tech.,
81—86,
Vol. 10.2478/pjct-2018-0042
20, No. 3, 2018 81

Batch electrodialysis of lactic acid obtained from LAB fermentation

Magdalena Lech*, Anna Trusek
Wroclaw University of Technology, Faculty of Chemistry, Division of Bioprocess and Biomedical Engineering, Wybrzeze
Wyspianskiego 27, 50-370 Wroclaw, Poland, e-mail: [Link]@[Link], [Link]@[Link]
*
Corresponding author: e-mail: [Link]@[Link]

The aim of this work was to develop the method of lactic acid (LA) separation from fermented whey. CMI-7000
Cation Exchange Membrane and AMI-7001 Anion Exchange Membrane were employed in electrodialysis process.
Experiments showed that the selected membranes separated organic acids effectively (including LA) from other
organic ingredients present in medium. Selecting an appropriate volume of a receiving chamber could lead to LA
concentration. Moreover, membrane fouling during separation was investigated. This phenomenon is negligible
which is the main advantage of this process. As it was shown during batch processes, with the voltage increase,
the rate of electrodialysis increases as well. It prompts to a reduction of residence time in electrodialyzer during
a continuous separation.

Keywords: lactic acid, membrane separation, Lactobacillus rhamnosus, whey, lactose.

INTRODUCTION cells. In effect, a purification of this stream is complicated

and difficult. Membrane reactor is increasingly popular
Lactic acid (LA) is one of the most common organic
process solution to produce organic acids13, 14. Its applica-
acids that is used widely in biotechnological and chemi-
tion leads to cells and heterogenic pollution separation.
cal technologies. For instance, in the food industry it
Much more complicated are the further steps. In fermen-
is widely used as an acidulant, a preservative and also
tation broth there are huge amounts of impurities such
as an emulgator1. In 2016 the yearly world production
as unreacted sugars, residue of proteins, or other organic
reached 1.077 · 106 tons and an annual growth rate
acids12. The most selectively method to separate these
14.2% is expected2.
compounds is an ion exchange chromatography14. Gon-
Beneficial properties of LA for skin condition have
zales et al.16 used two types of ion exchangers – Lewatit
been applied in cosmetics production. It is common
S2568H and Lewatit S3428 and obtained a final purity
used as an acidity regulator3 in cosmetics formulation,
of lactic acid higher than 99%. There is huge amount
but could be also used as active ingredients too. W. P.
of anion and cation exchange chromatography applied:
Smith4 treated patients’ skin with 12% w/v lactic acid
Reillex 42517, IRA-42018 or MWA–1 and AG3-X419. The
and this resulted in increased epidermal firmness and
ion exchange chromatography was an effective process
reduction of wrinkles amounts. LA after polymerization
in terms of the product quality, but it gave the product
as polylactide is a great alternative to botox5.
recovery in an unfavorable quantity approach.
A large application of LA comes with its consumption
LA can be also extracted by n-butanol. The advan-
for the synthesis of the polyester PLA (polylactic acid or
tage of this process is high selectivity of lactic, however
polylactide) – a biodegradable polymer. It is extensively
factors such as pH, mixing time, initial concentration
used in tissue engineering and in drug delivery systems6.
of lactic acid, and volume ratio between the organic
The global production capacity for PLA increases rapidly
and the aqueous phase affect the extraction yield20. An
every year and the capacity of 800.000 tons should be
integrated process including filtration, activated carbon
established in 20207 ([Link] Hence,
treatment and evaporation should be also mentioned20.
the lactic acid requirement is estimated at an annual
This method gives 85% concentration of LA. Another
growth of 5–8% per year8, 9.
integrated process of lactic acid recovery and purification
Not only PLA production consumes LA. This organic
process was esterification with distillation. Purified LA in
acid may be converted into a wide range of useful prod-
extraction process with n-butanol was mixed with metha-
ucts or semi - products such as acrylic acid, propylene
nol (in molar ratio 1:1.5) and small amount of sulfuric
glycol, 2,3-pentanedione, acetaldehyde, pyruvic acid1.
acid. The mixture was directed to fractionating column.
From the economical point of view and because of its
LA was a bottom fraction22. However, described process
enormous world consumption, new sources (especially
requires a lot of energy, therefore is not profitable.
natural – derived from food industry) used in lactic
This work tries to outline the promising field of LA
acid production are sought. It is quite obvious, that
purification by electrodialysis (ED). This approach is
a fermentation process is preferable than a chemical
intensively developing nowadays. Membrane processes
synthesis10. The fermentation is performed by lactic
are cheaper, more efficient than other purifying processes
acid bacteria – LAB11 that are able to convert simple
(e.g. chromatography or extraction) and works usually
carbohydrates such as glucose, galactose and lactose
in continuous system, what make them valuable from
(derived from whey) to LA. LA production from whey
industrial point of view. These are advantages and ED as
is certainly an eco-friendly process and gives surprising
a type of membrane process is characterized by the same.
fermentable capability12.
ED membranes are mechanically and chemically stable
A product of microbiological fermentation is contami-
in different electrolyte solutions23, so in effect they may
nated by unreacted substrate, residual of nutrient solution
be used for a long time, especially when it comes to
or – what is the most problematic – propagated bacterial
82 Pol. J. Chem. Tech., Vol. 20, No. 3, 2018

continues system processes. Additionally, regeneration nificant concentration) organic acids – especially citric
is quite easy and cheap (using NaOH, HCl or NaCl) and acetic acid. A presence of these acids generated
Many membrane systems have been already described a pH value about 4.0.
but this membrane technique is constantly studied. There In order to estimate a concentration of lactic acid salt
are many various types of this process: desalting elec- (during process outside acid there is formed its salt too)
trodialysis with membranes stacked alternately or water 1mL medium was mixed with 1M H2SO4 (Poch, Poland,
splitting electrodialysis with bipolar membrane, one stage CAS: 7664-93-9) and then lactic acid concentration was
system or multi – stage system etc.24, 25 measured with HPLC. The difference between based
This type of separation process is widely used to the on HPLC obtained value, and concentration (measured
separation and purification of organic acids26, besides in medium without sulfuric acid), is approximate salt
lactic acid, also citric acid27. Habova et al.28 received content.
a final concentration of LA (derived from a fermentation
broth) 151 g L–1. Boyaval et al.29 coupled electrodialysis Equipment
unit with continuous fermentation and achieved 85 g A laboratory equipment to electrodialysis (our con-
L−1 concentration in outlet stream. Kim and Moon25 struction) had three chambers divided by ion-exchanging
employed one-stage ED process, which lead to a high membranes (CMI-7000 Cation Exchange Membranes
volumetric productivity – 72 g L−1 h−1, that in compari- and AMI-7001 Anion Exchange Membranes, Membranes
son to a multi – stage separation leads to a decrease International, USA). Parameters of these membranes
in capital and operating costs. All of described above
processes refer to continuous process.
ED is very specific type of membrane process, because Table 1. The technical specifications of the ion-exchanging
as an one is a based on electric potential difference. membranes
Authors rarely investigate the problem of transport
through the membrane, which is undoubtedly very im-
portant from a process design point of view. They had
pretested results (often very satisfactory) of separation
made on industry or laboratory equipment, but do not
id not mention about mechanism.
In presented work, one – step batch electrodialysis pro-
cess was tested. An investigation a process using a batch
system could provide some information about mechanism
of separation, what is a quite difficult in continues sys-
tem case. CMI-7000 Cation Exchange Membrane and
AMI-7001 Anion Exchange Membrane from Membranes
International, USA, (which have not employed to LA
separation yet) were tested. These membranes should
have a great potential in LA purification30. Separated
fermented broth derived from membrane bioreactor
with Lactobacillus rhamnosus while post-production whey
was the carbon and energy source. The main task was are presented in Table 1. The area of each membrane
to separate LA from another small organic compounds was 3.2 cm2.
like lactose or peptides occurring in broth31. A cathode was placed in the left chamber (chamber
I, V = 30 mL) detached by the cation – exchanging
MATERIALS AND METHODS membrane while in the right chamber (chamber III, V
= 30 mL) an anode was placed. The electrodes were
Materials submerged in distilled water and connected with direct
current source (MS Major Science MP – 300V). A middle
Medium, which was purified, derived from membrane chamber (chamber II, V = 60 mL) was filed with a post
bioreactor with Lactobacillus rhamnosus strain immobi-
lized. In outlet stream there was microfiltration membrane
(pore diameter – 0.40 μm), so it leads to preliminary
purification of liquid from the largest structures and bac-
teria. Inlet stream of bioreactor consist of whey diluted
by the medium responded to the MRS32: (in g · L–1)
CH3COONa (5), Tween (1), K2HPO4 (2), Triamonium
citrate (2), MgSO4 · 7H2O (0.2), MnSO4 · 4H2O (0.05).
The whole process is precisely described in previous
work27.
Concentration of LA in outlet stream (simultaneously
in electrodialyzed solution) was approximately 7.2 g · L–1
(was not constant because came from microbiological
process). It contains small amount of unreacted lactose Figure 1. Scheme of an electrodialysis set
(1.4 g · L–1), proteins (0.5 g · L–1) and other (in insig-
 Pol. J. Chem. Tech., Vol. 20, No. 3, 2018 83

– culture medium. The scheme of the system described

is presented in Fig. 1.
During separation an electrical voltage, current,
temperature and the concentration of lactic acid were
measured.
Membranes were regenerated by 1% HCl (for the ca-
tion exchange membrane) and 1% NaOH (for the anion
exchange membrane) poured into an adequately chamber.
Additionally, ultrasounds for 5 min were applied.

Lactic acid analysis

The concentration of lactic acid was analyzed with
HPLC (Waters, USA) under isocratic conditions using
Synergii 4μ Hydro – RP 80 Å column (Phenomenex,
USA).
Samples were first filtered through a 0.22 μm syringe Figure 2. Organic acids present in whey (thin, pink line)
filter and then eluted with 20 mM K2HPO4/H3PO4 at and after fermentation process (thick black line):
pH 2.9 and 30°C for 10 min. The flow rate was 0.042 1st peak – lactic acid (retention time – 6.4 Min),
L h–1 and peak absorbance was monitored at 214 nm. 2nd peak – acetic acid (retention time – 7.5 Min ),
Lactic acid (Sigma – Aldrich, CAS: 79 – 33 – 4), citric 3rd peak – citric acid (retention time – 8.5 min)
acid (Poch, Poland, CAS: 538210118) and acetic acid
(Sigma – Aldrich, CAS: 64–19-7) were run as standards. L–1. Then the LA molecules, and also other organic
Under these conditions retention time for lactic acid acid molecules migrated to the right chamber using the
was 6.12 min and concentration was calculated using appropriate voltage – from 25 to 100 [V].
standard curve: At each voltage the concentration profiles in chamber
III were similar, but the process rate increased with
C [g L–1] = 4.35 ∙ 10–6 ∙ A (1)
a voltage increase (Fig. 3). After 5 hours of separation
where: A – area under area of peaks [μV·min] process, the concentration of LA was approximately 3.6
g L–1 (it estimated for 25% initial mass of LA). The
RESULTS AND DISCUSSION faster this condition was achieved the higher voltage was
The separation of LA from a fermented broth was
the main aim of this research. A processing stage of
LA synthesis in bioreactor is a complex microbiological
process, in which very various products can be formed.
Moreover, LA salts (sodium or calcium lactate) can
be found in fermented broth33. Electrodialysis process
does not distinguish these ingredients; lactate anions
derived from both acid and salts migrate to chamber
III. In effect at the equal volume of all chambers we
can obtain higher concentration of LA in chamber III
than in chamber II in the initial stage.
The experiment for total amount of LA determina-
tion (extraction by 1M sulfuric acid) indicated that in
our case almost 16% of LA molecules came from salts.
Figure 3. LA concentration in the right (III) chamber in time
Composition of separated medium of electrodialysis. The initial concentration in the
Electrodialysis is a specific membrane technique where middle (feed) chamber (II) C0,II_25V = 7.42 g L–1;
C0, II_50V = 7.43 g L–1; C0,II_75V = 7.25 g L–1;
a driving force is an electric potential difference. Dif-
C0 II_100V = 7.48 g L–1
ferent ions with the same sign migrate throughout the
membrane in the same direction. Thus, the presence of
applied. As expected, ions migrated after this time, but
other acids (other than LA) can be observed in chamber
with lower intensity.
III. They came from whey, which was used as a medium
During electrodialysis, the concentration of acids in
in bioreactor. L. rhamnosus is a homofermentative L-(+)-
the middle chamber decreased (Fig. 4).
lactic acid producer34; due to its metabolism only LA is
formed. Additionally, L. rhamnosus can used a citric acid Profile of resistance and temperature in the system
as well as lactose35, as a carbon source. Therefore, the
Electrical resistance and temperature were monitored
concentration of this acid decreases during fermentation
during the whole electrodialysis process. In each cham-
process (Fig. 2).
ber temperature was the same. For the first 20 minutes
Profile of product concentration in chamber III electrical resistance of the system decreased quite rapidly
and at the same time temperature increased for each
At the beginning, the middle (II) chamber was filled
applied voltage (Figure 5A and B). At the beginning of
with the fermented broth, microfiltrated previously.
the process the resistance of system is very high and the
Average LA concentration in this solution was 7.2 g
reason of this is a presence of distilled water in I and
84 Pol. J. Chem. Tech., Vol. 20, No. 3, 2018

and after initial time (in this case approximately 1 hour)

slowly decline is observed. A mentioned high temperature
at the beginning of separation is caused by I and III
chamber – there is very low concentration of ions - so
these chambers became resistors. As the concentration
increase the scheme does not put up such resistance
anymore. A further increase of temperature is present
because the response of this parameter is delayed.

Fouling on electrodialysis membranes

Figure 4. The change of LA concentration during electrodialysis
carried out at 25 V One of the disadvantages of any membrane separa-
tion is membrane fouling36. During electrodialysis not
III chamber (of course, an introduction of some ionic only acids can block the membrane surface. The post-
solution could decrease this phenomenon, but it could production fermented broth used as a feed in presented
lead to product contamination and that idea was aban- research contained unreacted lactose (approximately at
doned), so in effect the significant rise of temperature 1.5 g L–1) and whey proteins (approximately at 0.2 g L–1).
takes place. In order to check how the membranes are blocked,
After 1.5 hour of the electrodialysis process, tem- the experiment was performed with and without cleaning
perature was quite stable and LA concentration and procedure between successive periodic processes lasting
resistance increased modestly. These two last parameters 5 hours. As it is presented in Fig. 7 the process run with
are mutually bonded with each other. The higher acids the same efficiency on the regenerated membranes as
concentration in liquid in chamber III resulted their on the membranes using previously by 5 hours. It looks
lower concentration in liquid in chamber II. It leads to like using this type of membranes, fouling could be
the situation, in which there are less and less energy
conveyors and the middle chamber becomes a source
of resistance.
A relative change of temperature associated with
a given change in resistance for semiconductor (men-
tioned scheme is certainly one of them) is described by
equation:
[1]

where: Rt – resistance in given time, R0 – resistance at

the begging, ΔT – the difference between T and T0 ,
α – temperature coefficient. Figure 7. The profile of LA concentration in chamber III (the
Obtained results (profile of the temperature – Fig. 6A) process at 50 V) using the membranes with and
confirmed this theory, because according to it at the begin- without regeneration
ning of process there is rapidly increase of temperature

Figure 5. Profile of electrical resistance (A) and temperature (B) during electrodialysis separation

Figure 6. An example of the resistance, temperature and LA concentration in chamber III profile in time (electrodialysis at 100 V)
 Pol. J. Chem. Tech., Vol. 20, No. 3, 2018 85

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