Process Biochemistry 44 (2009) 353356

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Process Biochemistry
journal homepage: www.elsevier.com/locate/procbio

Short communication

Application of electrospun poly(ethylene terephthalate) nanober mat to apple

juice clarication
Beatriz Veleirinho, J.A. Lopes-da-Silva *
Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal

A R T I C L E I N F O

A B S T R A C T

Article history:
Received 8 March 2008
Received in revised form 25 October 2008
Accepted 5 November 2008

Electrospinning was used to produce self-supporting nanobrous poly(ethylene terephthalate)

membranes with good mechanical properties and straightforward handling. The application of this
type of membranes in apple juice clarication process was investigated. Processing characteristics and
quality parameters of apple juice were analyzed in order to compare the proposed method to traditional
clarication techniques. In general, the apple juice obtained from electrospun nanober membrane
ltration revealed physico-chemical characteristics comparable to those from juice claried by
ultraltration or by conventional clarication using ltering aids. Nevertheless, the new process showed
a high ux performance and revealed to be much faster, simple and more economical than the traditional
processes. This work demonstrated the ltration potential of an electrospun PET membrane thus
introducing a new concept of clarication and opening new approaches for the juice processing industry
or even for other food industry elds.
2008 Elsevier Ltd. All rights reserved.

Keywords:
Nanober mat
Electrospinning
Juice processing
Apple juice
Clarication
Membrane ltration

1. Introduction
Juice and fruit juice products represent a very important
segment of the total processed fruit industry and their consumption signicantly increased during the last years. The major
amount of apple juice is consumed as a brightly clear product
obtained through the clarication of the raw apple juice [1].
Traditionally, apple juice clarication is achieved by addition of
ltering aids, such as gelatin and bentonite, promoting the
adsorption and/or coagulation of a wide range of compounds.
The particles in suspension are then removed by centrifugation or
common ltration, improving juices stability and appearance and
thus increasing the consumers acceptability [2]. However, this
process is not only expensive and laborious but it can also causes
modications on sensory and nutritional properties of the juice
[3,4]. In addition, in recent years there has been an increasing
demand for natural, free-additive products, motivating the juice
processing industry to develop and employ free-additive clarication techniques. Ultraltration is the mostly widespread freeadditive clarication method due to many advantages, including
higher juice yield, cost reduction and high quality products [2,5].
Unfortunately, a disadvantage of this process is the rapid reduction
of permeate ux by fouling of the membrane which declines the
system performance [6].

* Corresponding author. Tel.: +351 234370360; fax: +351 234370084.

E-mail address: jals@ua.pt (J.A. Lopes-da-Silva).
1359-5113/$ see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.procbio.2008.11.008

Electrospinning technique has gained much attention due to its

ability to produce nanobrous mats with distinct characteristics,
such as high porosity, small pore sizes with an interconnected
structure and a large surface area per unit volume [7,8]. These
characteristics make them attractive in a variety of applications,
including both air and uid ltration [912]. In fact, in liquid
ltration applications, the nanobrous membranes produced by
electrospinning are expected to overcome some of the limitations
associated to the porous polymeric membranes manufactured by
conventional methods, e.g. low-ux and high-fouling performance,
due to their highly interconnected pore structure and large surface
area to volume ratio.
In despite of their potential as ltration membranes, to the best
of our knowledge the use of electrospun membranes in juice
processing has not yet been reported. Therefore, we have studied
the preparation and application of a nanobrous electrospun
ltration membrane for apple juice clarication. Poly(ethylene
terephthalate) (PET) was selected as the polymer to prepare the
nanobrous mats, due to its low cost, attractive structural and
mechanical characteristics, and good electrospinning properties
[13,14].
2. Experimental
2.1. Materials
The apples used in this study (Golden Delicious variety) were purchased from a
local market and washed with tap water to remove any adhering substances. PET
pellets used to produce the ltration membranes were kindly supplied by Flexitex
(Portugal). All solvents were analytical grade from SigmaAldrich Chemical
Company and were used without further purication.

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354

2.2. Preparation and characterization of the electrospun membrane

The nanobrous membranes were prepared by electrospinning 30% (w/v) PET
solutions in a mixture of triuoracetic acid (TFA) and dichloromethane (DCM)
(80:20 v/v), following the conditions previously reported [14].
Morphology and diameter of the nanobers were analyzed by scanning electron
microscopy (Philips XL 30 ESEM) at an accelerating voltage of 10 kV. The average
ber diameter was calculated from 300 measurements of the sample bers, using
the SEM images of magnication 1500 and appropriate software (Image J 1.37c,
Wayne Rasband, National Institute of Health, USA). The thickness of the electrospun
membranes was measured using a digital micrometer (model MDC-25S, Mitutoya
Corp., Japan). The apparent density and porosity of the brous membrane was
determined as previously described [13,14]. Mechanical properties were evaluated
by uniaxial tensile tests performed in parallel with the principal axis of ber partial
alignment using a texture analyser equipment (model TA.Hdi, SMS, England).
2.3. Preparation of apple juice
Peeled apples were introduced in a centrifugal juice extractor (Molynex
EazyClean) in order to obtain the raw apple juice. The juice was depectinized
with 0.75 mL/L Pectinex Ultra SPL (Sigma pectinase from Aspergillus aculeatus) and
0.15 g/L a-amylase (Fluka amylase from Bacillus subtilis, 49 U/mg), for 2 h at 50 8C.
The absence of detectable pectin and starch in the claried juice was conrmed by
ethanol and iodine method separately. The enzymes were inactivated by heat (90 8C
for 5 min) and the juice was ltered through a cheese cloth in order to remove the
remaining pulp before the clarication step.
2.4. Clarication of apple juice
Clarication of apple juice is normally performed by addition of clarication
agents (conventional clarication) or by ultraltration. For this reason, we have
used these two clarication methods as a reference to the ltration method using
the electrospun nanobrous mats proposed in this work. The unclaried juice was
used as a control for comparison among clarication treatments.
The conventional clarication method was based on a that previously described
by Gokmen et al. [2]. Accordingly, clarication was achieved by adding 0.5 g/L of
gelatin and 2.5 g/L of bentonite to the enzymatically treated juice, at 50 8C for 2 h.
The precipitate was removed by centrifugation (4000 rpm, 10 min) and the clear
juice was stored at 4 8C. The ultraltration technique was performed using a stirred
ultraltration cell (Millipore, model 8200). In order to produce the clear juice,
150 mL of enzymatically treated juice were ultraltered through a regenerated
cellulose membrane (Millipore, cut off 100 kDa), under N2 pressure. The claried
juice was stored until analysis at 4 8C.
Apple juice claried by electrospun nanobrous membrane (ENM) ltration was
obtained using also the stirred ultraltration cell (Millipore, model 8200). The ENM
was cut into a 63.4 mm diameter circle and introduced into the ultraltration cell.
The unclaried juice was forced to pass through the membrane using nitrogen
pressure. The claried juice was stored until analysis at 4 8C.
2.5. Juice characterization
Colour and turbidity were measured by UVvis spectrophotometry. Colour was
expressed as the extinction coefcient at 440 nm and turbidity as the percentage
transmission at 650 nm [2]. Soluble solids of samples were determined with a
manual refractometer (Atago, Tokyo), at 20 8C, and the results were expressed in
8Brix. The pH was measured using a pH meter (WTW InoLab). For titratable acidity

measurement, 25 mL of juice was titrated with 0.25 M NaOH to pH 8.1, and the
suitable volume was converted into malic acid equivalent. The total phenolic
content of samples was determined according to the Folin-Ciocalteau method [15].
Total protein was measured using a commercial protein kit (Micro Lowry,
Petersons ModicationSigma).
Free sugars (glucose, fructose and sucrose) where measured by HPLC using a
Beckman system with RI detector and an YMC-Pack polyamine II S-5um column
(250 mm  4.6 mm). Isocratic elution was carried out at room temperature using
acetonitrile/water (75:25) at 1.5 mL/min as the mobile phase. Organic acids were
quantied by HPLC using a Beckman System Gold, a diode array detector (210 nm)
and a Aminex Ion Exclusion HPX-87H column (300 mm  47.8 mm). Isocratic
elution was carried out at 40 8C using 5 mmol/L sulphuric acid at 0.6 mL/min as the
mobile phase.
2.6. Statistical analysis
A one-way ANOVA was used to test for any signicant difference between
treatments and control on each independent variable under study. A t-Student test
was performed to test for specic statistical signicance among data means.

3. Results and discussion

3.1. Membrane characterization
The electrospinning process was performed until the membrane reaches an average thickness of 0.20 mm (similar to the
ultraltration membrane used). The PET electrospun nanobrous
membrane (ENM) was typically characterized by a uniform
smooth surface and a good resistance to rupture despite of its
exceptional lightness. Fig. 1 shows the general aspect of the ENM
before and after apple juice ltration.
The electrospun membranes revealed good mechanical proprieties, such as a high tensile strength (2.7  0.2 MPa) and relatively
high elongation (35  8%) which are also important features for a
membrane ltration in order to avoid damage during handling. The
strength of the PET nanobers web was high enough to use for lter
without the need of any supporting matrix like meltblown or
spunbonded nonwovens, thus avoiding one of the common problems
of handling electrospun nanober mats, related to their usually low
mechanical strength.
The average density of the membrane (0.33 g cm 3) was 4.2
orders of magnitude lower than the original PET density which
reveals the presence of large amounts of pores among the bers. In
fact, the porosity of the ENM was about 80%.
Fig. 2 shows representative SEM images of the PET ENM,
revealing the detailed picture of the membrane morphology. This
analysis revealed a membrane with many attractive characteristics
for the required end-use as a ltration device: a random brous
arrangement, free of beads and with a tridimensional porous

Fig. 1. Images showing the general aspect of the PET ENM membrane (A) before ltration and (B) after apple juice ltration.

B. Veleirinho, J.A. Lopes-da-Silva / Process Biochemistry 44 (2009) 353356

355

Fig. 2. SEM images showing the morphology of the PET electrospun membrane (A) magnication 500; (B) magnication 1500.

structure, high porosity and small pore diameters. The average

ber diameter was 420 nm.
3.2. Clarication process
3.2.1. Processing characteristics
The most remarkable difference between the studied clarication processes was the total clarication time (Table 1). ENM
ltration was clearly the fastest clarication technique, being more
than 20 times faster than the conventional one. Additionally, heat
or clarication agents are not required by the ENM ltration
method, allowing a cost reduction, as well as avoiding off-avours
in the juice.
Comparing to the ultraltration technique the new proposed
method allowed for a faster juice ux during the process, thus also
providing less time consumption. An additional economical
advantage of the PET ENM ltration refers to a much lower work
pressure compared to the ultraltration process.
3.2.2. Changes on apple juice quality
The inuence of the different treatments on the juice quality is
shown in Table 2. All the treatments signicantly decreased the
apple juice colour (p < 0.05). The nal colour of the juice samples

was very similar for all treatments and visibly higher than the
control (unclaried juice). The preliminary removal of pectins and
starch, common to all the clarication methods, is the main
responsible for the improved colour and turbidity of the claried
juice samples. However, ENM ltration and ultraltration promoted a signicant decrease in the turbidity of the juice (p < 0.05)
and the best clarication efciency. No signicant effects of the
treatments were found for the total solids content, soluble solids,
pH and acidity (p < 0.05).
Table 3 shows the concentration of total phenolic compounds,
proteins, selected free sugars and organic acids. As expected, the
protein content of the apple juice is very low and relatively high
errors are associated to the results, which should be regarded as
indicative. Nevertheless, in general, a decrease in the protein
concentration was observed after the clarication treatments,
being more evident for the conventional clarication and PET ENM
ltration (p < 0.05). Since the protein content of the juice is very
low, removing this kind of compounds will not reduce the
nutritional value of this product, but can be useful to reduce
turbidity and improve juice stability.
In general, it was observed a more pronounced reduction of free
sugars when the clarication method was performed using the PET
ENM ltration. Under the analytical conditions used, no signicant

Table 1
Operational parameters for conventional clarication, ultraltration and PET ENM ltration, related to the treatment of 150 mL juice volume.
Method

Conventional clarication
Ultraltration
PET ENM ltration

Average processing
time (min)

Required
temperature (8C)

Required
pressure (psi)

Clarication
agents added

Average ux
(mL cm 2 min

160
35
6

50
N.A.
N.A.

N.A.
50.8
0.7

Bentonite and gelatin

N.A.
N.A.

N.A.
0.17
3.5

N.A. = not applicable.

Table 2
Physico-chemical properties (mean  standard deviation, n = 3) of apple juice samples obtained from different clarication treatments.
Treatment

Coloura Abs440 nm

Turbiditya% T650 nm

Total solids (%)b

Soluble solids (8Brix)b

Acidityb (%w/w)c

Unclaried (control)
Conventional clarication
Ultraltration
PET ENM ltration

0.53  0.07
0.41  0.04
0.40  0.03
0.40  0.03

75  4
82  4
87  2
88  1

13.1  0.6
13.2  0.6
13.0  0.2
13.1  0.7

14.5  0.4
14.5  0.5
14.2  0.2
14.4  0.5

0.33  0.03
0.32  0.03
0.35  0.01
0.32  0.02

a
b
b
b

a
b
c
c

Means with different letters within a column indicate signicant differences (p < 0.05).
All scores within each column were not signicantly different to p < 0.05 level.
c
As malic acid equivalent.
b

356

B. Veleirinho, J.A. Lopes-da-Silva / Process Biochemistry 44 (2009) 353356

Table 3
Composition in protein, phenolic compounds, sugars and organic acids (mean  standard deviation, n = 3) for apple juice samples obtained from different clarication
treatmentsa.
Unclaried (control)

Conventional clarication

Ultraltration

PET ENM ltration

Total protein content (mg L 1)

Total phenolic content (g L 1)b

194  16 a
0.15  0.01

83  16 b
0.13  0.01

145  14 c
0.14  0.02

98  3 b
0.15  0.02

Sugars (g/L)
Fructose
Glucose
Sucrose

95.4  0.8 a
37.2  0.3 a
13.6  0.5 a

91.3  0.5 b
35.2  0.5 b
12.3  0.3 b

94.8  0.9 a
33.7  0.8 c
13.1  0.4 a

89.9  0.6 b
33.6  0.4 c
10.9  0.4 c

Organic acids (g/L)b

Malic
Oxalic
Lactic
Citric

9.7  0.2
0.14  0.03
n.d.c
n.d.

9.9  0.3
0.10  0.02
n.d.
n.d.

9.8  0.1
0.18  0.05
n.d.
n.d.

9.8  0.2
0.13  0.03
n.d.
n.d.

Means with different letters within a row indicate signicant differences (p < 0.05).
Not signicantly different at p < 0.05 level.
c
n.d.not detected.
b

differences could be detected for the total phenolics and detectable

organic acids present in the different apple juice samples.
4. Conclusions
In this work, we have demonstrated that brous mats
composed of randomly oriented submicron-size bers can be
prepared by electrospinning and successful used, for the rst time,
as nonwoven lters in fruit juice clarication. Filtration using the
ENM allowed for a less time consuming and more economical
process, still originating a claried juice with characteristics
similar to that obtained by ultraltration.
It is important to point out that the investigated electrospun
membranes have not been fully optimized for further improvements. One relevant aspect, taking into account that the ndings
described came out from a lab-scale process, is the need for
further scale-up studies in order to understand how the process
would run on a larger scale. In addition, retention capability, ux
rate and the fouling problems can be further improved and
optimized by adequate control of the electrospinning process, in
order to manipulate ber diameter, porosity and membrane
thickness.
In addition, the possibility to incorporate bioactive compounds
or additional ber components into the nanobers may allow
preparing afnity membranes for the selective removal of certain
components, rather than operating merely by sieving mechanisms.
Acknowledgments
Authors gratefully acknowledge Fundacao para a Ciencia e
Tecnologia (FCT, Lisboa, Portugal) for funding through the
programmes POCI 2010 and FEDER (project POCI/CTM/58312/
04). The authors thank Dr. Manuel Rei (CITEVE, Portugal) for the
help with the SEM analysis. Authors also thank Mrs. Cristina Santos
(College of Biotechnology, Porto, Portugal) for helping with the
HPLC analysis.

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