Received: 5 May 2020 Revised: 27 July 2020 Accepted: 10 September 2020

DOI: 10.1002/app.50123

ARTICLE

Hierarchical amphiphilic high-efficiency oil–water

separation membranes from fermentation derived cellulose
and recycled polystyrene

Siju Cherikkattil Raghavan | Anju PV | Mudrika Khandelwal

Department of Materials Science and

Metallurgical Engineering, Indian
Institute of Technology Hyderabad,
Kandi, India
Correspondence
Mudrika Khandelwal, Department of
Materials Science and Metallurgical
Engineering, Indian Institute of
Technology Hyderabad, Kandi 502285,
India.
Email: mudrika@msme.iith.ac.in
Funding information
AT&T CSR
Abstract
A high-efficiency separation of oil and water can be achieved by using specially
designed amphiphilic porous membrane. However, the preparation of such
membranes often involves complex multistep chemical processes. Herein, we
report an amphiphilic composite membrane (polystyrene [PS]/bacterial cellu-
lose [BC] membrane) consisting of hydrophobic recycled PS and hydrophilic
BC, fabricated by a facile in situ fermentation process. Not only these mem-
branes exhibit a combination of contrasting wettability but also comprise of a
hierarchical network of microfibers and nanofibers, which makes them ideal
for oil–water separation. The structural and morphological properties of as-
produced BC, recycled PS membrane, and PS/BC composite membrane were
studied by Fourier transform infrared spectroscopy and scanning electron
microscopy, respectively. The ability of the membranes to separate oil and
water was tested by using an emulsion of hexane-in-water as the feed and the
collected filtrates were characterized by optical microscopy and UV–Vis spec-
troscopy. PS membranes were unable to separate oil and water, while the
PS/BC membrane efficiently separated water from the emulsion. PS/BC com-
posite membranes showed a high water recovery of more than 90%, against
only 57% recovery shown by BC. Mechanisms of oil–water separation for each
membrane are discussed. The reusability of the PS/BC composite membrane
was also demonstrated.

KEYWORDS
biopolymers and renewable polymers, cellulose and other wood products, membranes,
polystyrene

1 | INTRODUCTION demand for water due to rapid population growth, wors-

A large amount of wastewater, contaminated by oil is
produced every day from household activities, oil produc-
tion units, marine transportation, and industrial
processing, which poses a serious social and ecological
concern.1–3 The processing of wastewater is required to
allow reusability of wastewater, given the increasing
ening climatic conditions, and over-exploitation of water
resources. Many conventional technologies are applied
for the purification of oil-contaminated wastewater such
as centrifugation, gravity separation, ultrasonic separa-
tion, oil-absorbing materials, coagulation, flocculation,
and air flotation.4–9 However, due to various factors, such
as cost, large space requirement, low separation

J Appl Polym Sci. 2020;e50123. wileyonlinelibrary.com/journal/app © 2020 Wiley Periodicals LLC 1 of 8

https://doi.org/10.1002/app.50123
2 of 8
efficiency, and production of secondary pollutants, these
conventional methods for oil/water separation find lim-
ited application and require further research and
development.10–12
Out of the various oil–water separation techniques,
filtration membranes are preferred owing to relatively
simple operation, and better separation efficiency.13 Vari-
ous types of separation membranes with preferred and
tuned wettability- superhydrophobic and super-
oleophobic surfaces have been developed, where exam-
ples from nature such as lotus leaf and fish scales have
served as inspirations.14–21 However, both the types suffer
from specific disadvantages. In the case of super-
oleophobic surfaces, given that water is denser than most
oils, water would be deposited quickly on the membrane
surface and block the passage of oils. This lowers the
separation efficiency of oil removing membranes and
would require additional higher pressure, resulting in
high-energy consumption. On the other hand, in the case
of superhydrophobic membranes, these underwater
oleophilic membranes are inefficient when the oil is
heavier than water.22
Thus, there is an ongoing effort to develop
environment-friendly amphiphilic membranes for effi-
cient oil/water separation due to their differential wetta-
bility, high flux, and excellent selectivity.17,22–27 The
amphiphilic membrane can simultaneously act as super-
amphiphilic in air, oleophobic underwater, and hydro-
phobic under oil which makes them suitable for a
broader range of water–oil mixtures.
Cellulose, being the most abundant biopolymer, is
preferred owing to nonwoven fibrous morphology. Cellu-
lose fibers can be obtained from various sources includ-
ing plants, trees, tunicates, and microorganisms.28–30
Plant-based cellulose nanofibers have been explored for
the treatment of wastewater.31–33 However, the process to
obtain pure nanofibrous membranes from plants involves
several steps involving harsh and toxic chemicals and
high energy input.28–30 Therefore, although plant-based
cellulose is available in abundance, the overall conver-
sion and purification to obtain nanofibrous membranes is
bound to be more complex and less environment-
friendly. Further, the development of cellulose compos-
ites, to impart additional functionality, also requires
further chemical processes.
Bacterial cellulose (BC), produced by certain species
of bacteria (such as Acetobacter Xyllinum, Acetobacter
pasteurianum, Acetobacter rancens, Sarcina ventriculi), is
a three-dimensional porous network of 100% pure, semi-
crystalline cellulose nanofiber.34–36 The high wet strength
and nano-meso-microporous structure of BC makes it
one of the most suitable candidate for filtration
membranes.37–40 A few reports are available where BC,
RAGHAVAN ET AL.
being hydrophilic, has been chemically modified to
obtain an amphiphilic membrane. For instance, modifi-
cation of BC membrane by alkoxysilanes resulted in an
amphiphilic membrane and was successfully used to sep-
arate surfactant stabilized oil in water and water in oil
emulsions.40,41 In another work, functionalization of BC
with poly(fluorophenol) to tune the hydrophobicity and
durability was also reported.42 However, most of the
reported methods for the preparation of amphiphilic BC
membranes involve complex multistep chemical pro-
cesses and are not always environmentally friendly. Also,
the overall efforts to utilize BC or modified BC for the
intended purpose are limited.
Recently, our group reported a novel and facile envi-
ronmentally friendly method for producing amphiphilic
nano-microfibrous composite using in situ growth of
nanofibrous BC into hydrophobic microfibrous recycled
polystyrene (PS) membrane.43 The recycled PS mem-
brane was produced by a green, low cost, and low energy
process using orange peel extract by hand drawing
method.44 PS nanofibers based oil–water separation
membranes have been reported before for the separation
of diesel from water.45 Additionally, PS is used as a sor-
bent for oil spill cleanup, owing to its hydrophobic
nature.46 However, the utilization of recycled PS by prep-
aration of the amphiphilic membrane for oil–water sepa-
ration has not been reported before.
In this study, we present an in situ composite of BC
with recycled hydrophobic PS, which exhibits nano-
microfibrous architecture, as a media for hexane-water
separation. Industrially, n-hexane is used in many chemi-
cal processes involving the production of glues, leather,
textile, cleaning, degreasing, and oil extraction, petro-
leum refining.47 Thus, n-hexane has been used as a rele-
vant model for demonstrating the oil–water separation
ability of the material developed in this work. The amphi-
philic PS/BC membrane was prepared using a single-step
in situ fermentation process. To the best of our knowl-
edge, a PS/BC membrane with a combination of hydro-
philic nanofibers and hydrophobic microfibers for usage
in oil–water separation has never been reported.
2 | MATERIALS AND METHODS
Hestrin Schramm (HS) media used for the production of
BC includes glucose, peptone, yeast extract, citric acid,
and disodium hydrogen phosphate of microbiological
grade, which were purchased from the Himedia, India.
Recycled polystyrene membranes (PS) were obtained
from the Carbon Lab, Indian Institute of Technology,
Hyderabad. These membranes were produced by using
citrus peel extracts in which thermocol (made up of PS)
RAGHAVAN ET AL.
was dissolved to obtain a viscous solution that was hand-
drawn to produce nonwoven fibrous fabric.44 n-Hexane
(95% LR) was purchased from the S D Fine-Chem Lim-
ited, India. The surfactant, Tween 20 was purchased from
the Sigma-Aldrich, India.
2.1 | Preparation of amphiphilic PS/BC
membrane
BC membranes were produced by using the bacterial
strain ATCC 23769 cultured in HS media consisting of
glucose (20 g/L), peptone (5 g/L), yeast extract (5 g/L),
citric acid (1.15 g/L), disodium hydrogen phosphate
(3.4 g/L) and maintained at the pH between 4.5 and 5.0.
A 100 ml of HS media was poured in a petriplate and
inoculated with 3 ml of bacterial inoculum containing
3 × 108 colony forming unit (CFU)/ml. The composite
membranes of PS and BC were produced by in situ cul-
ture of BC in the PS membrane. For this, first PS mem-
branes were pretreated with ultraviolet radiation for 1 h
to sterilize, and then circular discs cut to fit petriplates
were placed in HS media.43 All culture plates were stati-
cally incubated at 26
C and harvested after 14 days. The
BC grows into the PS membrane during the culture and
results in a composite membrane referred to as PS/BC
membrane. BC and PS/BC membranes were repeatedly
washed using 0.5 N NaOH and water for removing media
components and bacteria. The wet pellicles were then
freeze-dried at −80
C under vacuum and used for further
analysis.
2.2 | Characterization
The surface morphology of the samples was observed by
field emission scanning electron microscope (FE-SEM)
(Model: SUPRA 40, Make: Carl Zeiss, Germany) at an
accelerating voltage of 5 kV. All samples were gold-
sputtered for about 10 s before imaging. Average fiber
diameter was obtained by considering atleast 100 fibers
in three different images. The functional group analysis
was done by Fourier transform infrared (FTIR) spectrom-
eter (Model: Alpha-P, Bruker Corporation) in the spectral
range of 400–4000 cm−1 in transmission mode. The
absorption properties of the feed and filtrates were ana-
lyzed by UV–Visible Spectrometer (Perkin Elmer, model
Lamda 365) in the range of visible wavelength range of
400–700 nm. The optical images of emulsified oil droplets
in the feed and the filtrates were observed by an optical
microscope (Leica DM2700M RL, Germany) in transmis-
sion mode, by placing a droplet placed between coverslip
and glass slide.
3 of 8
2.3 | Oil–water separation
The emulsified oil-in-water (o/w) mixture of 30:70 vol/
vol ratio of n-hexane to water was prepared and used as a
feed. About 0.5–1 wt% of Tween 20 was added as an
emulsifier in the above o/w mixture under vigorous stir-
ring at 1500 rpm for 8 h. The membranes (BC, PS, and
PS/BC) were prewetted with water and fixed between
two flanges of Buchner flask. About 25 ml of freshly pre-
pared homogenous emulsified o/w mixture was used as
an initial feed and poured onto the top of the membrane
across which a pressure difference of 200 mbar was
maintained. The volumes of filtrate collected for 25 ml of
feed through BC, recycled PS, and PS/BC membranes
were measured. The filtration experiment was repeated
in triplicate for each filter membranes and an average
value is reported here. The water separation efficiency or
recovery was calculated according to the following
equation:
η ð%Þ = V 2 =V 1 × 100
where V1 and V2 are the volumes of water in initial emul-
sified o/w mixture and the volume of water collected as
filtrate, respectively.
The reusability and water separation efficiency (%) of
PS/BC composite were studied for six times repeated
usage by thoroughly backwashing the used membrane
with 50% ethanol and DI water after each filtration.
3 | RESULTS A ND DISCUSSION
3.1 | Morphological and chemical
characterization
The surface morphology of BC, PS, and PS/BC mem-
branes were characterized using SEM and the micro-
graphs are shown in Figure 1. It is seen from Figure 1(a)
that the pure BC membrane comprises a porous three-
dimensional interconnected network of cellulose
nanofibers with an average diameter of 72 ± 15 nm. The
PS membranes comprise of fibers with diameter in the
range of 1–10 μm as shown in Figure 1(b). The PS/BC
composite is made of BC nanofibers interpenetrated
between the PS microfibers, which can be observed on
the surface of the composite (Figure 1(c)).The amount of
BC present in composites varied from 20% to 30% for the
given conditions of cellulose synthesis employed in
this work.
Further from SEM, it can also be seen that the PS
membrane possesses large pores of the order of several
microns while the BC membranes comprise of much
4 of 8 RAGHAVAN ET AL.

F I G U R E 1 Scanning electron microscopy images of (a) bacterial cellulose (BC), (b) polystyrene (PS), and (c) PS/BC membranes [Color
figure can be viewed at wileyonlinelibrary.com]

smaller pores (including micropores and mesopores).

Further, the composite membrane offers a wide distribu-
tion of pore size including micro, meso, and macropores.
The structural characterization of BC, PS, and PS/BC
membranes was performed by FTIR spectroscopy. The
FTIR spectrum of BC shows a broad absorption band at
3344 cm−1 due to the presence of OH group, peak at
2900 cm−1 for CH stretching, and a band between 1157
and 1053 cm−1 for C O C stretching (Figure 2(a)). The
absorption at 1426 cm−1 is attributed to the CH2 bend-
ing of primary alcohol in cellulose.48,49 The aromatic
C H stretching and methylene C H stretching vibra-
tion of PS are observed at 3020 and 2921 cm−1, respec-
tively, as shown in Figure 2(b). The aromatic C C
stretching is observed at 1433 cm−1 and C H bending is
noticed at 692 cm–1.46,50 For PS/BC composite, a combi-
nation of both BC and PS characteristic peaks are F I G U R E 2 Fourier transform infrared spectra of (a) bacterial
observed, which confirms the presence of BC and PS cellulose (BC), (b) polystyrene (PS), and (c) PS/BC composite
fibers in the composite.43 The presence of absorption [Color figure can be viewed at wileyonlinelibrary.com]
RAGHAVAN ET AL.
peaks at 3270 cm−1, and 1150–1023 cm−1 are mainly due
OH and C O C stretching from cellulose in the com-
posite. The peaks observed at 2918, 1643, and 1450 cm−1
are attributed to the methylene C H stretching, aromatic
C C stretching, aromatic C C stretching corr-
esponding to the presence of PS in the composite.
3.2 | Filtration studies
All membranes (BC, PS, and PS/BC) were prewetted with
water and then used as the filtering media for 30:70
emulsion of hexane and water. The filtrates were charac-
terized by optical microscopy as well as UV–Vis spectros-
copy, in addition to the visual examination. The
photographs and optical micrographs of feed and the fil-
trate are presented in Figure 3. The emulsion of oil and
water, that is the initial feed, is milky white in appear-
ance. The filtrates from BC and PS/BC composites mem-
branes show a noticeable difference from the feed and
are visually transparent as shown in Figure 3(b,d). The
filtrate from the PS membrane is milky white, similar to
the feed, indicating the inability of the PS membrane to
separate oil and water. It can be further seen from optical
micrographs, that feed contains oil droplets stabilized in
water, while filtrates obtained through PS/BC and BC
F I G U R E 3 Optical microscopy images and photographs of (a) feed, filtrate using (b) bacterial cellulose (BC), (c) polystyrene (PS), and
(d) PS/BC membrane [Color figure can be viewed at wileyonlinelibrary.com]
5 of 8
membranes do not show any presence of oil droplets.
The optical image of filtrate through the PS membrane
showed oil droplets similar to the initial feed, as expected
from the milky appearance of the filtrate. This shows that
the PS membrane does not separate oil and water and
thus, it allows the emulsion to pass through.
The transparency/turbidity of the feed and filtrates
was further studied by spectroscopy in the wavelength
range of 400–700 nm as shown in Figure 4. The feed
showed a high absorbance due to the presence of emulsi-
fied oil droplets. However, the filtrates of PS/BC mem-
brane and BC membranes, being transparent, did not
show any absorbance, which indicates the absence of
emulsified oil in the collected filtrates. As expected, the
filtrate of the PS membrane showed a higher absorbance
and only a slightly lower than that of the feed.
Overall, it is clear that BC and PS/BC membranes can
separate oil and water from the emulsion, whereas the PS
membrane is incapable of oil–water separation. The
mechanism of filtration for each of the membranes is dis-
cussed below.
The BC membranes comprise hydrophilic nanofibers
with a small pore size as seen from the SEM micro-
graphs.37 Upon, prewetting with water, BC membranes
provide a continuous path for water in the oil/water
emulsion to pass through. This retains the oil on the
6 of 8
membrane, which is further aided by small pore size, as
shown in Figure 5. Further, the mechanism of oil–water
separation for amphiphilic PS/BC membranes is also
F I G U R E 4 Absorbance spectra of feed and filtrates collected
through polystyrene (PS), bacterial cellulose (BC), and PS/BC
membranes [Color figure can be viewed at wileyonlinelibrary.com]
F I G U R E 5 Schematic diagram of oil–water separation through (a) polystyrene (PS) membrane, (b) bacterial cellulose (BC) membrane,
and (c) PS/BC membrane [Color figure can be viewed at wileyonlinelibrary.com]
RAGHAVAN ET AL.
schematically shown in Figure 5. During the first phase
of filtration, the water can easily pass through the PS/BC
membrane prewetted by water, due to the continuous
water phase and hydrophilicity of the membrane. As fil-
tration progresses and the volume of water decreases in
the feed, the oil droplets get closer leading to coalescence.
Finally, the oil phase forms a layer on the membrane sur-
face. This results in an efficient oil–water separation.
However, in the case of the PS membrane, due to the
large pore size and loosely connected fibers, the emulsion
passes through without any separation. Given, the capa-
bility of both BC and PS/BC membranes to separate oil
and water, the filtering efficiency is compared below.
The filtration data in terms of the amount of water
recovered, filtration efficiency are given in Table 1. It
should be noted that PS/BC membrane showed over 90%
water recovery while the BC membrane showed about
60% recovery. The lower efficiency of the BC membrane
may be due to the presence of its compact and
nanoporous network, which initially allows water pas-
sage but thereafter as the filtration progress, the pores are
blocked by coalesced oil droplets, which terminate the
separation process, as shown in Figure 5.
RAGHAVAN ET AL.
T A B L E 1 Filtration time and water separation efficiency of
BC and PS/BC composite filter membrane
Volume Volume Water separation
of feed of filtrate efficiency or
Membrane (ml) (ml) water recovery (%)
BC 25 10 ± 1 57
PS/BC 25 16 ± 1 90 ± 5
Abbreviations: BC, bacterial cellulose; PS, polystyrene.
F I G U R E 6 Separation efficiency of polystyrene/bacterial
cellulose membrane after repeated filtration [Color figure can be
viewed at wileyonlinelibrary.com]
Further, the transmembrane pressure was varied for
PS/BC membranes between 200 and 600 mbar. The fil-
trates were collected and tested by measuring absorbance
in the visible range. The filtrates collected at a working
pressure between 200 and 500 mbar showed the absence
of absorbance in the visible region with the water recov-
ery or water separation efficiency to be over 90%. At the
working pressure of 600 mbar, the PS/BC membrane was
unable to separate the water from the feed mixture. The
inefficiency of PS/BC membrane at high pressure may be
due to the physical damage of PS/BC membrane and also
the inability of oil droplets to coalesce together.
The reusability of PS/BC membrane was studied for
six times by backwashing with diluted ethanol and water
after each filtration run. The separation efficiency (%) of
the PS/BC membrane after each filtration was calculated
which showed over 90% water recovery for each run. The
collected filtrate was colorless and transparent and did
not show any absorbance in the visible region in UV–Vis
spectra as shown in Figure 6. The denaturation of PS due
to hexane prevented further usage.
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4 | CONCLUSION
A nano-microfibrous composite of amphiphilic PS/BC
membrane was successfully fabricated by in situ synthe-
sis of BC into recycled microfibrous membrane of PS for
oil–water separation. The unique combination of
oleophilic PS microfibers and hydrophilic BC nanofibers
provides contrasting wettability, along with continuity
and high porosity. The PS/BC membrane showed more
than 90% water separation efficiency (recovery) for six
cycles of usage. Pure BC also allows effective separation
of oil and water; however, the efficiency or recovery was
about 60%. The PS/BC membrane ensures a continuous
passage for water given the appropriate contrast in wetta-
bility, higher porosity, and pore size. This PS/BC mem-
brane produced by a combination of recycled PS and
renewable BC can be used as a promising filter media for
efficient water recovery from oily waste-water. The
design strategy reported in this work can be extended fur-
ther for the development of newer advanced materials.
ACKNOWLEDGMENTS
The authors would like to acknowledge Dr. Chandra
S. Sharma and his lab for providing PS fabric material.
The authors also acknowledge support from AT&T CSR
funds.
AUTHOR CONTRIBUTIONS
The manuscript was written through the contributions of
all authors. All authors have approved the final version
of the manuscript.
DATA AVAILABILITY STATEMENT
The raw/processed data required to reproduce these find-
ings cannot be shared at this time due to technical or
time limitations.
ORCID
Siju Cherikkattil Raghavan https://orcid.org/0000-
0003-2364-3028
Anju PV https://orcid.org/0000-0003-4758-1861
Mudrika Khandelwal https://orcid.org/0000-0001-9717-
5381
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How to cite this article: Raghavan SC, PV A,
Khandelwal M. Hierarchical amphiphilic high-
efficiency oil–water separation membranes from
fermentation derived cellulose and recycled
polystyrene. J Appl Polym Sci. 2020;e50123. https://
doi.org/10.1002/app.50123