Ultrasonics Sonochemistry 9 (2002) 225229

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Eect of ultrasound on the extractibility of corn

bran hemicelluloses
A. Ebringerov
a *, Z. Hrom
adkov
a
Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, 842 38 Bratislava, Slovakia
Received 9 July 2001; accepted 8 October 2001

Abstract
Various two-step extraction procedures with and without a short application of ultrasound in the rst step were used to examine
the eect of sonication on the extractibility and properties of the non-cellulose components of industrial corn bran (CB). The
polysaccharides recovered from the extracts of the rst (FI ) and second (FII ) extraction step were characterised by yield and composition. Using water as extractant in the rst step, similar yields of total extracted polysaccharides (FI FII ) were obtained by the
short sonication treatment when compared to the classical procedures using H2 O2 -containing alkaline media in the rst step. The
yields of FI FII showed a substantial increase (1040%) after application of ultrasound in 1% and 5% NaOH in comparison to
those of the classical procedures. The content of the neutral sugar components constituting the CB xylan varied from 65 to 88 mol%
in dependence on the conditions used in the rst extraction step.
2002 Elsevier Science B.V. All rights reserved.
Keywords: Zea mays L.; Corn bran; Extraction; Ultrasound; Hemicellulose; Xylan

1. Introduction
Corn bran (CB) represents a byproduct of the starch
production, available in huge amounts. It contains
at least 30% hemicellulose, comprising mostly a highly
branched heteroxylan type with properties similar to
those of exudate gums [1]. Although, no industrial applications of the corn bran xylan (CBX) have resulted
from past eorts, recently, several papers appeared [24]
dealing with the evaluation of novel and optimised extraction and purication processes of this potential
food gum. Usually, the extraction of CBX starts from
CB previously destarched by enzymic or ethanol-sieving
methods. Depending on the extractants used, the multistage procedures involve long extraction time and high
temperatures, as well as post-treatments to prepare highpurity products [3] what increase their costs. Extraction
of CBX from the unpuried CB with subsequent removal of co-extracted starch was presented as an alternative in our previous paper [5].

Recently, the application of ultrasound in extraction

and rening processes has found increasing attention [6
10]. Ultrasonication can improve signicantly the pectin
technology [11] and the industrial production of medicinal tinctures from herbs [12]. The eects of ultrasound on vegetal tissues during solvent extraction has
been described [13] and the ultrasonically assisted extraction of bioactive principles from herbs reviewed [14].
In our previous papers [5,15,16], the advantages of short
sonication treatments during the extraction of polysaccharides have been documented for various plant sources, such as corn cobs and various herbs. The objective
of this work was to examine the eect of ultrasound
on the extractibility of the non-cellulose components of
crude CB and the yield of isolated polysaccharides
(xylan-type hemicelluloses and starch) in comparison to
classical two-step extraction procedures.
2. Experimental
2.1. Material and general methods

Corresponding author. Tel.: +421-7378-2844; fax: +421-7-59410222.

E-mail address: chemebri@savba.sk (A. Ebringerova).

The CB (hulls) from Zea mays. L., a coproduct

generated by the starch production, was kindly provided

1350-4177/02/$ - see front matter 2002 Elsevier Science B.V. All rights reserved.
PII: S 1 3 5 0 - 4 1 7 7 ( 0 1 ) 0 0 1 2 4 - 9

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A. Ebringerova, Z. Hromadkova / Ultrasonics Sonochemistry 9 (2002) 225229

by Amylum Group plc, Slovamyl (Boleraz, Slovakia).

The air-dried bran was ground and sieved to yield a
material with particle size 0.25 mm. Ultrasonication
was carried out using the Ultragen system PERSON
(Nitra, Slovak Republic, 20 kHz) at sonic power of 100
W, sonication time intervals of 10 min, and ultrasonic
intensity 8 W cm1 . The generator was of the horn type.
The ground bran was dispersed at liquid to solid ratios
(L/S, ml/g) 50=2, 50=3, and 50=5, in distilled water, 1%
NaOH, and 5% NaOH. The dispersions were sonicated
for given time intervals in a glass beaker of standard
geometry in a water bath with controlled temperature.
The starch content of CB was determined by wetsieving using 70% ethanol [17]. The presence of starch in
the isolated polysaccharide fractions was checked by the
KI/I2 test giving a dark blue colour. Moisture content
was determined after drying samples to constant weight
at 105 C. The total carbohydrate content was determined according to the phenolH2 SO4 method [18] and
uronic acid content according to the 3-hydroxydiphenyl
assay [19]. The methods used for neutral and acidic
sugar analysis, and determination of protein have been
described in previous papers [5,17].

used with varying conditions in the I step, and constant

conditions (5% NaOH at room temperature for 1 h) in
the II step. All experiments were performed at least in
duplicate. Yields of the CBX preparations are given on
a dry weight basis related to the CB.
2.2.1. Classical procedures
In a typical experiment, the ground bran (50 g) was
suspended in 250 ml of the extractant heated to the given
temperature and stirred for 1 h. After rapid cooling to
room temperature, the insoluble material (residue RI )
was separated by centrifugation (20 min; 5000 g) and
subsequently treated (II step) with 200 ml of 5% NaOH
for 1 h at room temperature. The remaining insoluble
residue RII was separated by centrifugation as before.
The individual (EI , EII ) or combined extracts (EI EII )
were poured into four volumes of ethanol. The formed
precipitate was twice decanted with 80% ethanol, then
acidied with acetic acid to pH 6, followed by washing
with 80% ethanol, and then ltered o. The precipitate
was resuspended in distilled water and subjected to exhaustive dialysis in a cellophane bag. The non-dialysable
content of the bag was recovered by lyophilisation
yielding the CBX fractions FI , FII and FIII .

2.2. Corn bran xylan extraction and isolation

For the isolation of CBX, two-step alkaline extraction procedures, schematically illustrated in Fig. 1, were

2.2.2. Ultrasound-assisted procedures

In a typical experiment, the ground bran (5 g) was
suspended in 50 ml of the extractant and sonicated

Fig. 1. Scheme for the isolation of polysaccharide components of industrial CB by classical and ultrasound-assisted extraction procedures.

A. Ebringerova, Z. Hromadkova / Ultrasonics Sonochemistry 9 (2002) 225229

(sonic power 100 W) at the given temperature for 510

min. After separation of EI , the second extraction step
using 50 ml 5% NaOH as well as the treatments of
the supernatants from both extraction steps were performed at the same conditions as described in the classical procedures.

3. Results and discussion

Analysis of various CB preparations indicated considerable variations in the content of residual starch
depending on the particular milling operations used and
less dierences in the proportions of the non-starch
components (cell wall polysaccharides, protein, oil, phenolics, and ash) [24,20]. The residual starch content of
the industrial CB under investigation was 13.5% and the
neutral sugar content 66.5% (Table 1). The sugar analysis of CB revealed that xylose, arabinose and galactose
comprise 56% of the neutral sugars. Assuming that
these sugars and the uronic acids constitute the heteroxylan component of CB, then its deduced amount in
bran was about 39%, what is in accord with published
data of other bran materials [2,4,21].
A series of two-step extraction procedures (Fig. 1)
with and without application of ultrasound were used to
study the eect of sonication during the rst extraction step on the extractibility and recovery of the xylan
component of CB in comparison to various classical
methods. Sonication conditions (temperature and sonication time) were used at which degradation of CBX
molecules in aqueous alkali was reduced to a minimum,
what was documented in a previously reported study [22]
dealing with ultrasonic degradation of a well-dened
model CBX in various media.

Table 1
Analytical dataa of the CB
Ash
Protein (% N  6:25)
Starchb
Neutral sugarsc
Uronic acidsd

2.6
11.5
13.5
66.5
2.2

Neutral sugar componentse

Rhamnose
Arabinose
Xylose
Mannose
Glucose
Galactose

Tr
20.1
30.4
3.9
40.0
5.6

In % calculated on basis of absolutely dry CB.

Estimated by wet-sieving in 70% ethanol.
c
Determined by the phenolH2 SO4 method.
d
Determined by the 3-hydroxydiphenyl method.
e
Determined after acid hydrolysis by GLC as alditol triuoroacetates and expressed as relative wt.%.
b

227

The extractibility of CB and the yields of the isolated

polysaccharide fractions obtained by classical (AC)
and ultrasound-assisted (DF) procedures are listed in
Table 2. The yields of the isolated polysaccharide fractions from the classical procedures using 5% NaOH with
and without addition of hydrogen peroxide, and 10%
NH4 OH ranged between 35.2% and 40.6% (related to
the corn bran). The yields of the insoluble extraction
residues RII (2729%) are close to the cellulose content
of CB (2022%) [20] indicating a substantial removal
of the residual starch and hemicelluloses during extraction. However, only 4857% of the extracted material
(100%RII ) was recovered as polysaccharide fractions.
The rest is represented by dissolved lignin and low molecular compounds such as phenolic acids, and products from alkaline-oxidative degradation reactions of
the carbohydrate and lignin CB components.
The same mild extraction conditions (5% NaOH,
ambient temperature, 1 h), used for all procedures in the
second extraction step, enable the sonication eects to be
more evidenced. The eect of a short sonication treatment in the rst extraction step was studied in various
media, e.g. distilled water, 1% NaOH, and 5% NaOH.
As seen in Table 2, the yields of RII after sonication in
water varied between 33.1% and 30.1%, indicating that
the extractibility of bran was lower than in the case of the
classical extraction procedures. However, the amounts
of the totally isolated polysaccharide fractions achieved
a nearly same level. Although, the yield of residues RII
after application of ultrasound in aqueous alkali showed
only a slight increase compared to the classical procedures, substantially higher amounts of polysaccharides
were obtained due to their higher recovery from the alkaline extracts (6373%). This indicates that the carbohydrate components suer from degradation during the
ultrasound assisted extractions in a much lower extent
than during the classical procedures.
After sonication in 1% NaOH, the yields of polysaccharide fractions (FI FII ) increased to 44.450.9%
thus exceeding the corresponding yields obtained by the
classical extraction procedures by at least 10%. It is interesting that more than 70% of the extracted polysaccharides were recovered in the rst extraction step
(Table 2) which lasted only 510 min. A recovery of 78%
was found after the 10 min-sonication in 5% NaOH.
Under these conditions, the yield of FI FII rised up to
52.7% what is about 30% and 40% higher in comparison
to the yields obtained by similar extraction conditions
without application of ultrasound (method A) and by
the 5% NaOH/H2 O2 (method B), respectively.
The mechanochemical attack of ultrasound on bran
cell walls was signicant also in absence of alkali. Thus,
sonication in water at 60 C for 5 min (method D2) gave
similar yields of FI FII when compared to those obtained by the classical extractions using hydrogen peroxide-containing alkaline media.

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A. Ebringerova, Z. Hrom
adkova / Ultrasonics Sonochemistry 9 (2002) 225229

Table 2
Yieldsa of polysaccharide fractions (F) and extraction residues (RII ) obtained from CB by classical and ultrasound-assistedb extraction methods
Extraction method
Without ultrasound
A: 5% NaOH
B: 5% NaOH/H2 O2
C: 10% NH4 OH/H2 O2
With ultrasound
D1: H2 O
D2: H2 O
D3: H2 O
D4: H2 O
D5: H2 O
D6: H2 O
E1: 1% NaOH
E2: 1% NaOH
F: 5% NaOH

L/S

Extraction step I

Extraction step IIc

FI

T (C)

Time (min)

FII

RII

50/5
50/5
50/5

60
60
60

60
60
60

13.7

40.6d
35.2d
23.5

28.8
27.2
29.5

50/2
50/2
50/3
50/3
50/5
50/5
50/5
50/5
50/5

30
60
30
50
50
70
50
70
50

10
5
5
5
10
5
10
5
10

3.3
3.0
2.2
2.6
1.8
14.7
30.5
40.5
41.2

31.4
34.5
32.7
32.2
37.6
29.3
13.9
10.4
11.5

30.2
31.3
33.5
32.1
31.5
30.1
29.3
26.9
28.1

In % based on oven-dried CB.

Ultrasound power: 100 W.
c
Extraction step II: 5% NaOH, ambient temperature, 1 h.
d
Combined fractions FIII . L/S: Liquid to solid ratio.
b

The eect of L/S ratio, aecting the ultrasonic treatment, on the yield of isolated polysaccharides was examined for ultrasonication in water. A tendency towards
higher yields can be seen at lower L/S ratios (Table 2). In
all cases a positive temperature eect has been noted.
The obtained results unequivocally indicate a highly
positive eect of a short ultrasonical treatment on the
extractibility and recovery of polysaccharides from CB,
conrming similar eects observed in extraction processes of other plant materials [5,15,16].
It is to mention that the extractibility of CB is aected
by the dierent accessibility of the present residual starch
and cell wall polysaccharides. Whereas, the solubility of
starch can be enhanced by thermal treatment in water,

the xylan component needs the strongly concentrationdependent swelling action of dilute alkali and its ability
to cleave native bonds in the cell wall matrix. As seen in
Table 2, after the ultrasonical treatment in water at
70 C (method D6) the total yield of the isolated polysaccharide fractions increased substantially, particularly
that of FI (14.7%). Sugar analysis of the fraction (Table
3) revealed that it contains a considerable amount of
starch conrmed by the positive KI/I2 test and high
glucose content as well. As expected, variable proportions of co-extracted and co-precipitated starch were
present also in the other polysaccharide fractions. Based
on the analytical data (Table 3), the polysaccharide
fractions represent CB hemicelluloses, mainly of the

Table 3
Analytical data of polysaccharide (CBX) fractions isolated from CB by classical and ultrasound-assisted extraction methods
Method

Fraction

Without ultrasound
A
B
With ultrasound
D5
FI
FII
D6
FI
FII
E1
FI
FII
F
FI
FII

N (%)

Neutral sugar composition (mol%)

UA (%)

Ara

Xyl

Man

Glc

Gal

1.2
0.9

20.6
15.8

31.1
27.9

Tr
1.1

45.3
50.3

3.2
4.9

2.9
2.1

2.2
1.1
2.3
1.8
1.6
2.7
1.4
0.3

25.7
26.7
18.9
32.0
18.0
21.4
21.1
20.4

13.5
40.1
25.7
49.9
21.2
30.4
30.8
37.5

0
0
0
Tr
0
0
1.1
2.1

60.8
30.7
55.4
11.9
55.7
45.2
41.6
34.3

0
2.5
0
5.2
5.1
3.0
5.4
5.7

1.5
3.3
nd
3.8
2.9
2.1
3.1
1.6

UA: Uronic acids determined by the 3-hydroxydiphenyl method.

A. Ebringerova, Z. Hromadkova / Ultrasonics Sonochemistry 9 (2002) 225229

xylan-type (CBX), contaminated with starch and coextracted proteins. The constituting sugars of the xylan
component of CB, i.e. xylose, arabinose, and galactose
[2,17] comprise 6588% of the neutral sugars in polysaccharide fractions from the second extraction step
(FII ). These fractions are less contaminated by starch
when compared to those obtained by the classical procedures. The results indicate that the short ultrasonic
pretreatment may be used as rening step in the preparation of CBX. However, contaminating proteins and
starch can be removed also by enzymic post-treatments.

4. Conclusions
The eciency of the ultrasound-assisted two-step
extraction procedures exceeded that of the classical
procedures depending on the extractant used in step I.
Similar yields of total extracted polysaccharides can be
achieved by sonication in water, whereas, the yields were
signicantly higher using aqueous NaOH solutions. The
whole extraction time can be reduced due to the shorter
extraction time of step I with the high-temperature
regime. These advantages support the importance and
great potential of ultrasound-assisted extraction processes applied for the isolation of industrially important
polysaccharides from plant materials.

Acknowledgements
The authors acknowledge funding by the Slovak
grant agency VEGA (grant no. 2/7138) and the Slovak
Academy of Sciences (COST D10/016/01).

229

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