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Fabrication and characterization of

cellulose acetate/hydroxyapatite composite

membranes for the solute separations in

Aneela Hayder, Arshad Hussain, Ahmad

Nawaz Khan & Hizba Waheed

Polymer Bulletin

ISSN 0170-0839
Volume 75
Number 3

Polym. Bull. (2018) 75:1197-1210

DOI 10.1007/s00289-017-2084-1

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Fabrication and characterization of cellulose acetate/

hydroxyapatite composite membranes for the solute
separations in Hemodialysis

Aneela Hayder1 • Arshad Hussain1 • Ahmad Nawaz Khan1 •

Hizba Waheed1

Received: 10 March 2017 / Revised: 12 May 2017 / Accepted: 31 May 2017 /

Published online: 8 June 2017
Ó Springer-Verlag GmbH Germany 2017

Abstract Asymmetric composite membranes of cellulose acetate (CA)/hydroxya-

patite (HA) are prepared using phase inversion method for the separation of water,
urea, glucose, and protein. Various concentrations of HA up to 15 wt% are dispersed
uniformly in the CA matrix. The pore size of the CA matrix is tunable by varying
the concentration of HA. The flux of water, urea and glucose is *7 times higher
while for BSA *12 times higher in CA/HA composite membranes than pure CA.
Similarly, water absorption capacity of CA/HA composite membranes are increased
owing to the incorporation of hydrophilic HA in CA matrix. Moreover, the retention
rates are also reasonable in CA/HA composite membranes depending on the dif-
ference in porosity of the membranes. The permeation of urea is much higher in the
CA/HA composite membranes as compared to pure CA, which is more suitable for
hemodialysis. Thereby, such CA/HA composite membrane offers the potential to be
used in hemodialysis applications.

Keywords Composite membrane  Cellulose acetate  Hydroxyapatite 

Hemodialysis  Permeation

& Arshad Hussain

& Ahmad Nawaz Khan
School of Chemical and Materials Engineering, National University of Sciences and
Technology, Sector H-12, Islamabad, Pakistan

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Dialysis technology is gaining importance for the kidney treatments and became a
million dollar industry since the past 20 years. Thousand of people are under
hemodialysis and the mortality rate is estimated to grow at 18% annually at a
constant rate. The major challenge in the developing countries is not only the
availability of treatment, but also the affordability of the service that should be
addressed to improve outcomes for patients with kidney failure diseases.
Hemodialysis is the most economical and safe purification technology, which has
been assessed with better outcomes among other treatments such as peritoneal
dialysis and kidney transplantation [1, 2]. The principle of hemodialysis is to
remove the accumulated uremic toxins, excess water and ions from the body of
patient and purification of blood takes place by membrane technology [3]. The
advent of membrane technology made not only the process of purification feasible,
but also playing a vital role for the removal or recovery of toxic or valuable
components from the waste products.
In hemodialysis, the blood and dialysate are kept separated within the dialyser by
a semi-permeable membrane that removes the undesirable products from the body
of the patient for blood purification [4]. Solute clearances are completely depending
on the patient’s condition. To improve the solutes clearances, various strategies are
applied during hemodialysis including more efficient dialyzer, increasing the time
of dialysis and optimizing the blood and dialysate flows during each dialysis
treatment [5, 6]. However, there are variations in values of urea, glucose and other
solute removals in achieving therapeutic goals depending on the type and quality of
membranes. Various methods are used for the manufacturing of the membranes like
sol–gel method, sintering, track, etc., however, phase inversion method is most
widely used due to its ease and well established understanding based on the
thermodynamic as well as kinetic theories of polymer solution [7, 8]. In phase
inversion, asymmetric membranes are prepared by immersing the casting solution
film in a non-solvent coagulation bath. As a result of diffusional exchange among
solvent and non-solvent, phase separation takes place for forming the asymmetric
structure of the membrane [9, 10].
Among the wide range of polymers, only few polymers like polyimide, ethylene
vinyl alcohol, poly(methyl methacrylate), polyacrylonitrile and cellulose acetate
(CA), etc., are used as membrane materials owing to their physical and chemical
characteristics [10–12]. Particularly for dialysis membrane, CA is the most widely
investigated system due to its renewable, biodegradable and biocompatible nature as
well as low cost and good toughness as compared to other biopolymers [13, 14].
Early investigations reported novel CA membrane blended with phospholipid for
hemocompatible filtration in which resulted membrane exhibited excellent blood
compatibility such as protein adsorption resistivity as compared to neat CA
membrane [14, 15]. Moreover, CA/PEG casting membranes were prepared by
varying the concentration of PEG, the pore size increased to 50 lm and also
increased the permeability of the membrane reported by Saljoughi et al. [16]. Kee
and Idris fabricated the hemodialysis membranes using different molecular weight

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of CA with the varying concentrations of monosodium glutamate (MSG) from 2 to 6

wt%, which enhanced the permeability performance of uremic toxins [17].
Although various additives and nanofillers are incorporated into CA matrix,
however, still the challenge exists to achieve the remarkable efficiency in improving
the permeability and selectivity of the membranes for absorbing the harmful
Hydroxyapatite (HA) has attracted much attention as a biomolecules carrier
among other nanofillers due to its excellent bioactivity and biocompatibility. It has
good adsorption to protein and is mainly used for repairing of bone tissue in the
medical field because of its biocompatibility, bioactivity and osteoconductivity
[6, 18]. For many years, HA has been used in adsorption chromatography and
usually applied for separating various proteins as a column in a high performance
liquid chromatography apparatus [19]. It has been known that HA has two different
binding sites on its surface, i.e., C sites are rich in calcium ions or positive charge
which bind to acidic groups of proteins and the P sites attach to basic groups of
proteins due to lack of positive charge [20].
In this research work, the composite membranes of CA with hydroxyapatite (HA)
nanofiller are prepared using the phase inversion method. The fluxes as well as
permeability of the composite membranes are investigated using bovine serum
albumin (BSA) as a model protein. By the addition of HA up to 10 wt%, the
macropores are formed in CA/HA composite membrane whereas with further
increase of HA up to 15 wt%, the pore size is decreased which affects eventually the
permeability of the membrane. Such CA/HA composite membranes are useful to
separation and permeation of water, urea and glucose.

Experimental section


Cellulose acetate (CA) (Mw = 30,000) was used as a matrix of the polymeric
membrane purchased by Sigma-Aldrich, USA. Food grade formic acid (FA) with an
analytical purity of 98% (Merck) was used as a solvent and distilled water was used
as the non-solvent agent. PEG 400 was used as pore former purchased by Sigma-
Aldrich, USA. Solute experiments were performed with glucose, urea and bovine
serum albumin (BSA) solutions. Hydroxyapatite (HA) nanoparticles were synthe-
sized in the laboratory and dispersed into the CA matrix at different concentrations
of 5, 10 and 15-wt% and thereby termed as CA/HA-5, CA/HA-10 and CA/HA-15,

Synthesis of hydroxyapatite (HA)

For the synthesis of HA nanoparticles, the method described in our previous work
[21] was followed in which the sources of calcium and phosphorus were combined
using calcium nitrate tetrahydrate (Ca(NO3)24H2O) and diammonium hydrogen
phosphate ((NH4)2HPO4) by adjusting the pH at 12 using ammonia solution. Firstly,

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the solution of Ca (NO3)24H2O was prepared by dissolving 130 g of its quantity in

800 ml distilled water at 80 °C along with continuous stirring. Then using the same
conditions, the solution of (NH4)2HPO4 was prepared by adding 38 g in 550 ml
distilled water. The pH was maintained at 12 by the addition of ammonia solution.
Both solutions were mixed by the drop wise addition of Ca(NO3)24H2O solution
into (NH4)2HPO4 solution along with stirring for 2 h at 80 °C. The solution was
then left for 24 h for the precipitation of HA followed by repeated washing with
distilled water. After filtration, the powder was dried at 80 °C for 24 h and calcined
at 550 °C for 3 h. The standard diffraction data cards used for HA was JCPDS
09-0432 and matched the appearance of the Bragg’s reflections as well as correlated
with the XRD patterns reported by Ain et al. [22]. Moreover, the spherical shape of
the HA nanoparticles were formed and the size of the particles was in the range of
60 ± 20 nm [23].

Preparation of CA/HA composite membranes

Membrane was prepared by phase inversion method with variation of HA in CA as

shown in Table 1. Formic acid (FA) is used as a solvent and required amount of HA
is dispersed into FA which is stirred for 5 h and sonicated for 30 min. In the
meantime, CA and PEG 400 (used as a pore former) were dissolved in FA. The
weighed amounts of both solutions; HA and CA/PEG solutions were mixed together
and kept on stirring for 24 h. The casting solution is then casted on the glass plate
with the help of doctor blade of 200 lm thickness. The glass plate was then
immersed in the distilled water bath at room temperature and post treated with hot
water of about 90 °C so that evaporation of solvent could take place at higher rate to
obtain the good porosity. Afterwards, the membranes were dried in the oven at
50 °C for 24 h. The thickness of the composite membranes was varied from 200 to
250 lm.

Characterization of composite membranes

Equilibrium hydration

The equilibrium hydration determines the water sorption capacities of a material for
the transfer of cell nutrients and metabolites and also for the good absorption of
body fluids. The water sorption capacities are determined by weighing the dry
sample and then soaked the membrane in distilled water for 24 h at room

Table 1 Compositions of
Samples CA (wt%) PEG (wt%) FA (wt%) HA (wt%)
different CA/HA composite
Pure CA 20 4.6 75.4 0
CA/HA-5 20 4.6 70.4 5
CA/HA-10 20 4.6 65.4 10
CA/HA-15 20 4.6 60.4 15

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temperature and weighed after mopping with blotting papers. The percentage of
water sorption from the two samples was obtained by:
ESW ¼ Wwet Wdry =Wdry  100% ð1Þ
where, ESW is the percentage water sorption of CA/HA membranes at equilibrium,
Wdry is the weight of the dry membranes after the drying in vacuum oven at 50 °C
and Wwet indicates the membrane weight at equilibrium water sorption.

Flux measurements

The pure water and solute flux of the membranes are measured at 0.8 bar. Water
flux was calculated as per the following expression;
Jw ¼ Q=ADt ð2Þ
where, Jw is the flux (L/h m2), Q is the amount of permeate collected (L), Dt is the
sampling time (h), A is the effective surface area (m2) of the membrane.

Membrane resistance (Rm)

One of the important parameter is the resistance to the membranes for different
pressures. Pure water flux is measured at different transmembrane pressures like 20,
30, 40, 50, 60 and 70 kPa. From the slope of the transmembrane pressure difference
(DP) vs. water flux (Jw) plot, resistance of the membrane was evaluated as per the
Rm ¼ DP=Jw : ð3Þ

Separation studies

The performance of CA/HA composite membranes in terms of rejection of BSA

(Sigma-Aldrich 60 kDa), urea and glucose are evaluated using the permeation cell.
Solutes with the concentration of 1 mg/ml were selected for the testings. The
experiments are carried out at room temperature at 0.8 bar. The percentage of solute
rejection was determined using the following expression,

%R ¼ 1  Cp =Cf  100% ð4Þ
Cp is the concentration of permeate (mg/ml), Cf is the concentration of feed (mg/
ml). The concentrations of BSA, urea and glucose in permeate and their absorbance
is calculated by UV–visible spectrophotometer at wavelength of 280, 520 and
700 nm, respectively.

Permeation studies

The permeation characteristic of the pure CA and CA/HA composite membranes

(having thickness of approximately 0.2–0.25 mm) for three different solutes, i.e.,

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urea, glucose and bovine serum albumin (BSA) are studied at room temperature.
The ultrafiltration nalgene cell is used for the experiment having a cylindrical
hollow chamber. The known concentrations of the solutes are introduced on the feed
side and then by determining the flux per unit pressure drop per unit membrane
thickness gives the permeation values of different solutes. The calculation is done as
per the expression,
P ¼ J=DP  l ð5Þ
where, P is the permeability (L/h m bar), J is the flux of the solutes (L/h m2), DP is
the pressure difference and l is the thickness of the wet membrane (m).

Scanning electron microscope (SEM)

The morphology of the samples is determined using a scanning electron microscopy

(SEM) (JEOL-instrument JSM-6490A). The samples are frozen in liquid nitrogen
where it becomes fragile and broken to generate the fresh surface. The samples are
mounted on aluminum stubs and coated the surfaces with gold.

Fourier transform infrared spectroscopy (FTIR)

Infrared (IR) absorption spectra are measured on a Perkin-Elmer SPECTRUM-1000

Fourier transform IR (FTIR) spectrophotometer with resolution of 1 cm-1, in the
range of 4000–450 cm-1 using KBr pellets.

Results and discussion

In this study, different compositions of hydroxyapatite (HA) in cellulose acetate

(CA) matrix are prepared. Moreover, the effect of HA on the morphology, pure
water flux, urea clearance, glucose and BSA retention, membrane resistance, water
sorption capacity, separation and permeation is discussed.

Morphology of CA/HA composite membranes

To observe the porosity of the membranes, SEM morphology of pure CA and CA/
HA composites is shown in Fig. 1. The pure CA (Fig. 1a) is showing smooth texture
without any visibility of macro pores. The average pore size in pure CA is
*30 ± 3 nm. Whereas, the macro pores are formed in CA matrix (Fig. 1b, c) by
the incorporation of HA up to 10 wt%. The average pore size is relatively small
*72 ± 5 nm in CA/HA-5 and becomes larger *115 ± 10 nm in CA/HA-10.
Moreover, with the addition of HA up to 15 wt% (Fig. 1d), apparently the macro
pores are not visible and pore size is reduced to 50 ± 5 nm. Moreover, HA
nanoparticles are also agglomerated in the CA matrix as relatively large particles are
visible in the image. Evidently, HA concentration plays a vital role to optimize the
pore size of the CA matrix, which would eventually influence the performance of

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Fig. 1 Morphology of a Pure CA, b CA/HA-5, c CA/HA-10, and d CA/HA-15

the membranes. Noticeably, the reduction in average pore size at 15 wt% HA is

possibly due to difference in the viscosity of the solution. Usually, up to 10 wt%
CA/HA composite membranes, the viscosity of solution is not high due to which
HA nanoparticles can leach out at higher rate, leaving macro pores in the CA
matrix. However, at 15 wt% HA, the viscosity of the solution is relatively high so
that HA could not leach out at high rate for the formation of macro pores and
thereby, the pore size remained smaller. The porous morphology is desirable as
porous matrix permits the solutes to pass through more readily as compared to
compact/dense matrix.
In Summary, the incorporation of HA in CA showed not only good interfacial
interaction, but also uniform dispersion as viewed by SEM. There is no visible
agglomeration of HA nanoparticles up to 10 wt% in the CA matrix.

Interaction of CA/HA composite membranes

Figure 2 shows the FTIR result of pristine HA, pure CA and CA/HA-10 composites
in the range of 450–1500 cm-1 to highlight their interaction. The spectrum of
pristine HA is consistent as reported earlier by Zavastin et al. [24] in which the
appearance of peaks at 474, 564, 603, 963 and 1044 cm-1 corresponds to the
various modes of PO43- groups in HA. In comparison to the FTIR spectra of pure
CA and CA/HA-10, the intensity of peak appearing at 603 cm-1 increases while the
peak of HA at 564 shifts to 558 cm-1. The increase in the peak intensity reflects

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Fig. 2 FTIR spectrum of Pure CA, pure HA and CA/HA-10

higher ordering of HA crystals in the CA matrix. Moreover, the HA peak at

1044 cm -1 disappears in the composites. The disappearance as well as slight
shifting of the HA peaks in the composites clearly indicate the interfacial interaction
among CA and HA owing to the consumption of the functional groups involved in
the formation of the bonds among CA and HA.

Flux measurement of CA/HA composite membranes

The hydrophilic nature of PEG and HA also act as a driving force for water sorption
in the membrane along with the membrane porosity. Figure 3 shows the effect of
composition on the flux of water, urea, glucose and BSA. Pure CA exhibits higher
flux value for water *30 L/h m2 and intermediate flux values for urea and glucose
which are *21 and 13 L/h m2, respectively. While lower flux is obtained for bovine
serum albumin (BSA) having the value of *2 L/h m2. Sivakumar et al. [25]
mentioned the flux of pure CA for water *26.8 l/h m2 which is close to our
measured value. In case of CA/HA composites, the values of flux increased
significantly as compared to the neat CA. At concentration of up to 10% HA, fluxes
are increased to 198, 142, 98 and 36 L/h m2 for water, urea, glucose and BSA,
respectively. Overall the increase in flux of water, urea and glucose is *7 times
higher while for BSA *12 times higher in CA/HA composite membranes than in
pure CA. However, by further increase in concentration to 15% HA, values of fluxes
have been reduced relatively. The increase in the flux up to 10 wt% HA in CA
matrix is owing to the larger pore sizes of the membranes. The decline in flux
beyond 10% HA is due to the reduction in pore size. The closed segmental
arrangements of the CA/HA-15 sample may be due to agglomeration of HA in the
polymer chains and formed a less or no voids between them and forms a denser

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Fig. 3 Fluxes of water, urea, glucose and BSA in pure CA, CA/HA-5, CA/HA-10, and CA/HA-15

structure in the sub layer which does not create an empty space between polymer
chains for the formation of pores in the membrane. Moreover, the pore size of the
composite membranes is tunable by increasing the HA concentration which would
eventually affect their fluxes and permeabilities as investigated and discussed
further [26, 27].

Membrane resistance

Resistance of the membrane is determined by applying various pressures to the

membranes and taking the inverse slope of the linear relationship between pressure
versus flux, the value of Rm is calculated and presented in Table 2. Pure CA has the
highest value of Rm *2.02 kPa/L h-1 m-2 as compared to composite membranes.
The value of Rm decreases to 1.82 and 1.76 kPa/L h-1 m-2 for 5 and 10% HA in CA
and then increase to 1.89 kPa/L h-1 m-2 for 15% CA/HA composites. Conse-
quently, increase in HA concentration to 10% increases the pore size and reduces
the resistance towards pressure. At higher HA concentrations (above 10 wt%),
porosity has been decreased and dense structure is formed thereby increasing the Rm
value relatively.

Table 2 Membrane resistances

Samples Rm (kPa/L m-2 h-1) Water absorption capacity (%)
and water absorption capacities
for different CA/HA Composite
Pure CA 2.02 46
CHA-5 1.82 52.6
CHA-10 1.79 56.7
CHA-15 1.89 54

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Water absorption capacities

The relationship between different concentrations of HA and equilibrium water

uptake is shown in Table 2. The percentage of water uptake by pure CA membrane,
when hydrated for 24 h in ultrapure water is 46%. In case of CA/HA-5 and CA/HA-
10 membranes, water absorption percentage is increased to 52.6 and 56.7%,
respectively. Moreover, CA/HA-15 also exhibit higher water uptake *54%. The
high equilibrium water uptake observed in these membranes is probably due to more
hydroxyl groups in the composite membranes.

Separation measurements of CA/HA composite membranes

Figure 4 shows the percentage retention of the BSA, glucose and urea solutions for
different CA/HA composite membranes. Pure CA membrane has a BSA rejection
rate of 98% while urea and glucose *45 and 58%, respectively. 98% BSA rejection
for pure CA was reported by Idris et al. [28]. By the addition of HA in CA,
membrane performance has been improved in terms of urea and glucose clearances.
The percentage retention for both urea and glucose decreases at concentration of
10% HA, to 32 and 45%, respectively. It means that 68% of urea and 55% of
glucose is permeating or clearing out through the membrane while BSA retention
for this CA/HA-10 membrane is 89%, which is also reasonable retention rate.
However, increasing the concentration beyond 10% of HA, increases the BSA, urea
and glucose retention to 93, 42 and 50%, respectively, owing to the difference in
their pore size.
Retention rates may also be affecting due to the difference in the molecular
weight as the molecular weight of urea is lower than glucose and BSA. However,

Fig. 4 Retention of urea, glucose and BSA in pure CA, CA/HA-5, CA/HA-10 and CA/HA-15

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this may also be due to the porosity of the CA/HA composite membranes. As the
pore size is largely reduced in CA/HA-15 composites, which causes the higher BSA
retention. In case of CA/HA-10, there is more open porous structure, which may be
due to the fact that leaching rate of HA is much higher at lower concentrations of
HA as compared to the higher concentrations during gelation process and thus
retention of BSA is reduced relatively.

Permeation measurements of CA/HA composite membranes

It is believed that permeability depends basically on the hydrophilicity, porosity and

thickness of the membranes. These characteristics enhance the permeation
properties and membrane performance. Thin selective layer always have the ability
of high flux and high permeability properties, because the exterior layer governs the
membrane transport [29]. The permeability of all membranes has been calculated
using Eq. 5 and shown in Fig. 5.
In case of water and urea permeation, pure CA shows less permeation of about
0.005 and 0.0037 L/h m bar, respectively. By the addition of HA to the
concentration of 10 wt%, permeability of water and urea reached to 0.075–0.035
L/h m bar, respectively, as shown in Fig. 5. As compared to the pure CA, urea
permeability has been increased to about 10 times for 10 wt% HA. Porosity and
hydrophilicity of these CA/HA-5 and CA/HA-10 membranes may be the one reason
for higher permeation rate. CA/HA-15 showed a marginal decrease in the urea
permeability (i.e., 0.021 L/h m bar) owing to lesser pore size.
The glucose permeability for pure CA and other composite membranes is less as
compared to urea. This may be due to the fact that glucose has higher molecular
radius (5.5 Å) as compared to the smaller radii of urea (1.8 Å). CA/HA-10 exhibits

Fig. 5 Permeability of urea, glucose and BSA in pure CA, CA/HA-5, CA/HA-10 and CA/HA-15

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the maximum glucose permeability of 0.018 l/h m bar which is decreased when HA
concentration reaches to 15 wt% (0.01 l/h m bar).
In case of BSA permeability, pure CA is permeable to BSA, but in a very less
quantity of 0.0001 L/h m bar which increases to 0.0059 L/h m bar for CA/HA-10.
CA/HA composite membranes permits only trace amount of BSA as compared to
the urea and glucose. For the application of hemodialysis, the uremic toxins like
urea should be cleared out and the substances, which are necessary for the patients,
should be prevented [30]. BSA is used as a standard for essential proteins, the
permeation of which may compromise the nutritional status of the patient. Such
permeation amount of BSA is well within the allowed range of albumin leakage.
However, a modest loss of albumin does not cause a risk to patients. Normally
albumin loss can be up to 30–35% [31, 32], but in this research work, it is only 11%
in more porous membrane. So far, it can be depicted from the studies that composite
membranes are hydrophilic and permeability of low molecular weight solutes is
superior to the pure CA, but showing some selectivity towards urea. However, these
composite membranes also permit less amount of BSA protein to pass through them,
which is a very important characteristic of hemodialysis membranes. It is also
reported that HA and CA have very good cytocompatibility and blood compatibility
which promote their suitability towards hemodialysis applications [33].


CA/HA composite membranes are synthesized by varying the concentrations of HA

using the phase inversion method. Fluxes for pure water, urea, glucose and BSA are
investigated. Results from this study indicate that composite membranes have good
uremic toxin permeabilities and reasonable BSA retention. In summary, these
membranes also exhibit better surface roughness and good hydrophilicity as
compared to the pure CA membrane. Since the use of HA and CA membranes for
dialysis application proved to be promising. Biocompatibility test on the membranes
will be performed in the future. CA/HA-10 membrane has good performance in
terms of excess water, urea and glucose with higher permeability and good water
sorption capacities as compared to other fabricated membranes.

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