LWT - Food Science and Technology 59 (2014) 486e494

Contents lists available at ScienceDirect

LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Physical quality and in vitro starch digestibility of bread as affected by

addition of extracted malva nut gum
Anchalee Srichamroen*
Department of Agro-Industry, Faculty of Agriculture, Natural Resources and Environment, Naresuan University, Phitsanulok Province 65000, Thailand

a r t i c l e i n f o a b s t r a c t

Article history: Malva nut gum (MNG) was extracted by alkaline solution. The previous studies showed that the extract
Received 28 June 2012 had viscosity and gelling properties as well as inhibited a-amylase activity in starch solution 1.5e2.0
Received in revised form folds higher than that of original malva nut gum. This research was aimed to investigate the a-amylase
14 April 2014
inhibitory effect of alkaline-extracted MNG in solid food and to determine physical properties of MNG-
Accepted 24 April 2014
containing bread. The scanning electron microscopy of in vitro digestibility with a-amylase of MNG-
Available online 4 May 2014
containing breads showed less porosity and more undigested starch granules remained intact with
the matrix compared to control. This finding was consistent with the reduction of glucose (33e40%) and
Keywords:
Malva nut gum
maltose (23e39%) levels compared to that of control after a-amylase digestion for 180 min in a dialysis
Bread system. The results showed that extracted MNG significantly (p < 0.05) increased loaf volume, and
Digestibility moisture content by 1.5e12%, and 8.2e12.8%, respectively compared to that of control. Addition of
Dialysis extracted MNG in bread formulation significantly reduced moisture loss and firmness of the bread crumb
Scanning electron microscopy after storage for three days.
Ó 2014 Elsevier Ltd. All rights reserved.

1. Introduction containing galactomannan compared to wheat bread without gal-

Increased postprandial plasma glucose during 2 h after meal of
type 2 diabetic subjects (T2D) is associated with the incidence of
cardiovascular disease (Ceriello, 2005; Heine & Dekker, 2002). A
clinical goal of treating diabetic subjects is to decrease postprandial
hyperglycemia and cardiovascular risk factors. Diets play a role in
preventing the rapid rise of plasma glucose levels in the post-
prandial state. Viscous fibers have been well-known to reduce
postprandial blood glucose concentrations in humans and animals
(Anderson, Akanji, & Randles, 2001). It was proposed that the fully
hydrated chains of soluble fibers diminish the contact between
glucose molecules and small intestinal mucosal cells therefore
reduce the rate of digestion and absorption of carbohydrate
(Blackburn, Redfern, & Jarjis, 1984; Rainbird, Low, & Zebrowska,
1984; Torsdottir, Alpsten, Anderson, & Einansson, 1989).
Addition of hydrocolloid to enhance functional properties of
bread has been investigated by increasing number of researchers
(Woolnough, Monro, Brennan, & Bird, 2008). These authors re-
ported a lower rate of in vitro starch digestibility of bread
* Tel.: þ66 55 962746; fax: þ66 55 962703.
E-mail address: anchalee1@gmail.com.
actomannan (Brennan, Blake, Ellis, & Schofield, 1996; Slaughter,
Ellis, Jackson, & Butterworth, 2002). Other studies used guar gum,
locust bean gum, and xanthan gum reported increasing the weight
of baked products, improving dough development (Rosell, Rojas, &
Benedito, 2001), increasing gas retention of dough, improving
texture of crumb and crust by controlling moisture retention
(Huttner & Arendt, 2010). However, these results are not consistent
in some studies. Addition of b-glucan in bread formulation
contributed to a reduced loaf volume and increased crumb firmness
compared to control wheat bread (Gaosong & Vasanthan, 2000;
Gill, Vasanthan, Ooraikul, & Rossnagel, 2002; Symons & Brennan,
2004). Addition of hydroxypropylmethylcellulose, a hydrocolloid
that forms thermoreversible gel networks in baked products,
increased crumb firmness when added at levels higher than 1.5 kg/
100 kg starcheflour blend basis (Sabanis & Tzia, 2011).
Malva nut fruit (Scaphium scaphigerum (G. Don) Guib. et Planch)
is native to the South East Asia and China. In Thailand, people have
used the mucilaginous substance of the seeds as traditional medi-
cine for laxative benefits. My laboratory has succeeded in produc-
ing dietary fibers extracted from malva nut seeds. The extract
contains total dietary fiber 80 g/100 g, protein 2 g/100 g, ash 7 g/
100 g (Srichamroen & Chavasit, 2011a). Malva nut gum (MNG)
extracted with alkaline solution had carboxylic bonds with the

http://dx.doi.org/10.1016/j.lwt.2014.04.046
0023-6438/Ó 2014 Elsevier Ltd. All rights reserved.
 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494
reduction of galacturonic acid contents. This is correlated with the
increased storage moduli (G0 ) of alkaline-extracted gum with the
value being higher than that of water-extracted MNG. The previous
study also showed the a-amylase inhibitory effect of alkaline-
extracted MNG in starch solution (Srichamroen & Chavasit,
2011b). The alkaline-extracted MNG is new to scientific report. It
is interesting to investigate the ability of MNG to inhibit a-amylase
activity in solid matrix and to determine the effect of extracted
MNG on physical properties of bread.
2. Materials and methods
2.1. Materials
Malva nut seeds harvested during March and April in 2010 were
obtained from local markets in the East of Thailand. Malva nut gum
was extracted in the laboratory at Naresuan University
(Srichamroen & Chavasit, 2011a). Briefly, the seeds were ground
(0.5 mm mesh size) and dispersed in distilled water at a ratio of
1:100 (w/v) and placed in a boiling water bath for 1.5 h. The slurry
was then cooled to room temperature, and added with NaOH to a
final concentration of 0.05, 0.1, or 0.2 mol/L, then treated with ab-
solute ethanol at a ratio of 1:1 (v/v). Precipitate was removed to
recover MNG. The gum was adjusted to pH 7.4 and dried in hot air
oven at 60  C. All of extracted MNGs were ground to pass through a
100 mm sieve before use.
All chemicals, pepsin and a-amylase used in this study were of
analytical grade and purchased from Sigma Chemical Co. (St. Louis
MO, USA). Dialysis membrane (molecular weight cutoff of 12,000)
was obtained from Membrane Filtration Products Inc. (Seguin
Texas, USA).
2.2. Bread preparation
Wheat bread, called control bread, consisted of 60 g wheat flour,
1.5 g dried yeast, 3.5 g soybean oil, 0.5 g salt, 0.5 g sugar, and 34 g
water. Because of high water binding capacity of extracted MNG
(Srichamroen & Chavasit, 2011a), water level of MNG-containing
bread formulation had to be adjusted in order to avoid underde-
veloped gluten network due to increased dietary fiber content.
Previous research showed that MNG-containing bread should have
an additional 6 g of water when MNG was used 1.7 g/100 g flour.
Extracted MNG was boiled to fully hydrate and cooled to room
temperature prior to be added into the mixture of bread formula-
tion. Dough was fermented at 40  C and 90% relative humidity for
2 h in the chamber of incubator (ShelLab model TC2323, Cornelius
OR, USA), after which they were kneaded and divided into 210 g
portions and put in a bakery stainless steel tin (19 cm length, 8 cm
width, 8 cm height). Breads were baked in an air oven (Kluay Nam
Thai Trading Group Co., Ltd., Bangkok, Thailand) at 170  C for
20 min. After baking, breads were cooled at room temperature for
1 h before analyses. For the study of effect of storage on physical
properties of bread, bread loaf was kept in a polypropylene bag at
4  C to prevent mold. After certain time of storage (on days 1, 2 and
3) bread loaf in a food-grade polypropylene bag (Goldenpack Co.,
Chonburi, Thailand) was transferred to room temperature before
further analyses. The reason to choose three days of storage was to
imitate the use of bread in a local hospital store for diabetic
patients.
2.3. Loaf volume and specific volume
Loaf volume was determined by rapeseed displacement ac-
cording to AACC approved Method 10-05 (AACC, 2000). Loaf height
was determined using calibrated calipers which measured from the
487
center of the loaf. Specific volume was calculated by dividing loaf
volume by weight.
2.4. Moisture content and water activity
Moisture content of bread crumb was determined by heating
samples in an oven at 105  C for 12 h. Three bread slices were cut
from the central loaf and a crumb was taken from the centre of each
slice and torn into small pieces for moisture determination. Water
activity of bread crumb was measured at 25  C using a Novasina
AWC200 water activity meter (Axair AG, Pfäffikon, Switzerland).
2.5. Crumb firmness
The crumb firmness measurement was performed with a QTS
Brookfield Texture Analyser (Middleboro, MA, USA). Three bread
slices were cut from the central loaf. The central part of each slice
was cut into cubes (2  2  2 cm) and subjected to one compression
test. The sample was compressed to 40% original height at a
compression load of 25 kg with a cross-head speed for 60 mm/min.
2.6. Scanning electron microscopy (SEM)
Bread crumb was cut into fine pieces with a blade in order to
create a clean fracture surface to observe in the scanning electron
microscope (LEO model 1455 VP, LEO Electron Microscopy Ltd.,
Cambridge, England). Sample was mounted on an aluminum
sample holder (12 mm diameter) with double sided conductive
carbon tape and a line of carbon paint was painted around the base
of the sample to improve conductivity from the top of the seed to
the taped surface. The sample was then sputter coated with gold
(Sputter Coater Model SC 7620, Quorum Technologies Ltd., UK) and
placed in the SEM chamber for examination and photographed
using a 10 kV of electron beam-accelerating voltage.
2.7. In vitro digestibility of starch of the breads
To simulate the hydrolysis reaction in human stomach, pepsin
(5.75 units/g starch, Sigma Chemical Co., St. Louis MO, USA) was
added into 1 mol/L sodium phosphate buffer solution (pH 6.9)
which contained fine pieces of bread equivalent to 40 g of wheat
flour (Symons & Brennan, 2004). The mixture was adjusted to pH
1.5 (with HCl) at 37  C for 30 min. The pH of mixture was readjusted
to pH 6.9 (with NaOH) prior to addition of porcine pancreatic a-
amylase (110 units/g starch, Sigma Chemical Co., St. Louis MO, USA).
The mixture was transferred to a dialysis bag against 250 mL of
deionized water in erlenmeyer flask, and stirred at 37  C in a water
bath shaker. Dialysate (2 mL of deionized water in erlenmeyer flask)
was collected after 15, 30, 60, 120, and 180 min in order to deter-
mine glucose and maltose contents.
2.8. Sugar determination
Glucose and maltose contents in the dialysate were measured by
using HPLC system (Shimadzu, Kyoto, Japan). The dialysate (2 mL)
was filtered through a 0.45 mm filter and a portion (10 mL) of filtrate
was injected into the HPLC column (InertsilÒ NH2 5 mm, dimension
4.6  250 mm). The mobile phase was CH3CN/H2O:75/25 at flow
rate 1.0 mL/min, and 40  C. The HPLC was performed using a
refractive index detector with detection limit of 0.01 mmol/L.
2.9. Statistical analysis
All experiments were performed in three replications. Results
were presented as the mean  SE. Statistical analysis was
488 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494

performed by using SPSS version 15.0 for window. Repeated Mea-
surement ANOVA was used for sequenced periodical studies, fol-
lowed by StudenteNewmaneKeuls (SNK) test as a post-hoc
analysis to detect any significant differences among the groups. A p-
value < 0.05 was considered statistical significance. For interval
periodical study, one way ANOVA was used followed by SNK at p-
value < 0.05.
3. Results
3.1. General appearance
MNG-containing breads showed qualitatively large and small
open crumb cells more than that of control (Fig. 1). Bread con-
taining 0.5 g/100 g MNG showed mixtures of big and small gas cells.

Fig. 1. Slices of control and MNG-containing breads. (A) control bread; (B) bread containing 0.5 g/100 g MNG extracted with 0.05 mol/L NaOH; (C) bread containing 1 g/100 g MNG
extracted with 0.05 mol/L NaOH; (D) bread containing 0.5 g/100 g MNG extracted with 0.1 mol/L NaOH; (E) bread containing 1 g/100 g MNG extracted with 0.1 mol/L NaOH; (F)
bread containing 0.5 g/100 g MNG extracted with 0.2 mol/L NaOH; (G) bread containing 1 g/100 g MNG extracted with 0.2 mol/L NaOH.
 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494
Table 1
Physical properties of control and MNG-containing breads.
Bread Height (cm) Loaf volume
(mL)
Control 7.73  0.03a 700  2.90g
MNG extracted with 0.05 mol/L NaOH
0.5 g/100 g concentration 6.53  0.08d 770  1.15b
1.0 g/100 g concentration 7.03  0.14c 742  1.53d
MNG extracted with 0.1 mol/L NaOH
0.5 g/100 g concentration 6.60  0.02d 720  2.89e
1.0 g/100 g concentration 7.40  0.05b 710  1.73f
MNG extracted with 0.2 mol/L NaOH
0.5 g/100 g concentration 6.76  0.06d 785  2.89a
1.0 g/100 g concentration 7.06  0.03c 750  2.77c
Data present as mean  S.E. of three independent replicates.
aeg
different letters following means within the same column indicate a significant
difference at the p < 0.05 level.
While 1 g/100 g MNG containing bread showed big gas cells with
fewer amounts of small gas cells.
MNG-containing breads had significantly lower height than did
control (Table 1). Addition of 0.5 g/100 g MNG in bread formulation
decreased the height of baked bread by 12e15% of control. While
the reduction in bread height was 4e9% with the addition of 1 g/
100 g MNGs in bread formulation.
3.2. Loaf volume and specific volume
Addition of 0.5 g/100 g alkaline-extracted MNGs in bread
formulation increased loaf volume by 3e12% of control, while
incorporation of 1 g/100 g MNG in the bread resulted in 1.5e7%
increase in loaf volume compared to the control (Table 1). MNG-
containing breads had significantly higher specific volume
compared to that of control, except for bread containing MNGs
extracted with 0.1 mol/L NaOH which had the lowest loaf volume
among MNG-containing breads (Table 1). Specific volume involves
both loaf weight and loaf volume. Weights of MNG-containing
breads were not much different, thus loaf volume was a factor
affecting on specific volume. The low specific volume of 0.1 mol/L
NaOH extracted MNG-containing bread might be explained by the
different peak of spectral FT-IR pattern at 898 and 823 cm1 (CeH
and CeC bond, respectively) which was intensely found in 0.1 mol/L
NaOH extracted MNG (Srichamroen & Chavasit, 2011a). I could
speculate that MNG extracted with 0.1 mol/L NaOH had high water
holding capacity which led to slightly disturb expansion of gluten
network and reduced loaf volume.
Table 2
Moisture contents of control and MNG-containing breads during three day storage.
Bread Moisture content (g/100 g)
Day 0
Control 40.44  0.86b
MNG extracted with 0.05 mol/L NaOH
0.5 g/100 g concentration 43.78  0.41a
1.0 g/100 g concentration 45.61  0.31a
MNG extracted with 0.1 mol/L NaOH
0.5 g/100 g concentration 43.98  0.56a
1.0 g/100 g concentration 45.47  0.29a
MNG extracted with 0.2 mol/L NaOH
0.5 g/100 g concentration 41.81  0.42b
1.0 g/100 g concentration 44.39  0.31a
Data present as mean  S.E. of three independent replicates.
aee
different letters following means within the same column indicate a significant difference at the p < 0.05 level.
489
3.3. Moisture content and water activity of baked breads
Specific volume Addition of extracted MNGs in bread formulation significantly
(mL/g) increased moisture content by 8.2e12.8% of control bread, except
3.80  0.06b for bread containing 0.5 g/100 g MNG extracted with 0.2 mol/L
NaOH which increased moisture content by 3.4% of control
4.20  0.06a (Table 2).
4.13  0.03a
After 24 h of storage at 4  C, the moisture content of control
3.60  0.06c bread was lost by 6%, whereas moisture content of MNG-containing
3.56  0.09c breads was lost by 1.6e6.0% (Table 2). The moisture content of
control bread on day 2 was 9.4% lower than that of day 0. While the
4.20  0.01a
moisture content of all MNG-containing breads on day 2 ranged
4.16  0.03a
between 3.0 and 7.3% lower than that of day 0. After 3 days of
storage, the control had significantly lowest moisture content
compared to MNG-containing breads.
The water activity of control bread at day 0 was 0.946, while
MNG-containing breads had water activity level ranging from 0.950
to 0.964 (data not shown). After 3 days of storage, the water activity
of control was 0.937. Water activity values of MNG-containing
breads did not significantly change from day 0 of each formula-
tion. These results were in agreement with that of breads con-
taining other hydrocolloids which had little change of water
activity between fresh and older crumb (Abu-Ghoush et al., 2008;
Czuchajowska, Pomeranz, & Jeffers, 1989).
3.4. Crumb firmness
Fresh MNG-containing bread significantly increased crumb
firmness by 13e16% of control (Fig. 2). Although fresh MNG-
containing bread had higher firmness than that of control, the
increased rate of firmness after storage of MNG-containing bread
was much lower than that of control. The low rate of increased
firmness of MNG-containing breads was associated with decreased
loss of moisture content.
After 24 h of storage at 4  C, the firmness of control bread was
increased by three folds, while the firmness of MNG-containing
breads was increased by 1.05e1.5 folds (Fig. 2). The firmness of
control bread on day 2 and day 3 of storage was much increased
more than that of MNG-containing breads. The slope coefficient of
increased firmness of control bread was 8.2 (r ¼ 0.92). While the
firmness of all MNG-containing breads on day 2 and day 3 of
storage was gradually increased with slope coefficient ranged be-
tween 2.2 and 3.2 for 0.5 g/100 g MNG-containing breads and 2.7e
3.9 for 1 g/100 g MNG-containing MNG breads (r ¼ 0.85e0.95). At
day one and beyond all MNG-containing breads had softer crumb
than did control bread. Specifically, crumb firmness of control was
1.6e2.1 folds higher than that of MNG-containing breads.
Day 1 Day 2 Day 3
37.86  0.18e 36.62  0.31c 36.43  0.22c
42.00  0.17bc 42.48  0.27a 41.93  0.23a
42.96  0.08a 42.27  0.14a 42.16  0.16a
41.50  0.28cd 41.25  0.30b 41.52  0.31a
42.50  0.28ab 41.00  0.23b 41.40  0.26a
41.10  0.21d 40.61  0.18b 40.11  0.22b
42.30  0.17ab 41.24  0.14b 41.57  0.29a
490 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494

Fig. 2. Changes in crumb firmness of control bread and MNG-containing breads during three day storage. Symbols: control; 0.5 g/100 g of 0.05 M NaOH MNG;
0.5 g/100 g of 0.1 M NaOH MNG; 0.5 g/100 g of 0.2 M NaOH MNG; 1 g/100 g of 0.05 M NaOH MNG; 1 g/100 g of 0.1 M NaOH MNG;
1 g/100 g of 0.2 M NaOH MNG. Data present as mean of three independent replicates.

Table 3
Maltose contents in dialysate (mmol/L) of in vitro digestion in a bread e a-amylase-dietary fiber system.

Bread Maltose content in dialysate (mmol/L)

15 min 30 min 60 min 120 min 180 min

a a a a
Control 4.1  0.05 4.3  0.13 4.8  0.10 4.9  0.05 5.3  0.15a
MNG extracted with 0.05 mol/L NaOH
0.5 g/100 g concentration 2.6  0.03b 2.8  0.05b 2.9  0.10c 3.1  0.03d 3.3  0.09c
1.0 g/100 g concentration 2.5  0.07b 2.6  0.03b 2.8  0.07c 3.0  0.01d 3.2  0.07c
MNG extracted with 0.1 mol/L NaOH
0.5 g/100 g concentration 2.6  0.01b 2.7  0.10b 3.0  0.15bc 3.5  0.07bc 3.7  0.05b
1.0 g/100 g concentration 2.5  0.05b 2.7  0.05b 2.9  0.10bc 3.4  0.01c 3.6  0.01b
MNG extracted with 0.2 mol/L NaOH
0.5 g/100 g concentration 2.7  0.07b 2.8  0.05b 3.3  0.13b 3.7  0.18b 4.0  0.05b
1.0 g/100 g concentration 2.5  0.13b 2.7  0.03b 3.0  0.10bc 3.5  0.15bc 3.8  0.18b

Data present as mean  S.E. of three independent replicates.

aed
different letters following means within the same column indicate a significant difference at the p < 0.05 level.

3.5. In vitro digestibility
Before food reaches the small intestine, enzymatic digestion
with pepsin and acidic condition in the stomach are factors
effecting the structure of food matrices. To simulate the condition
of gastrointestinal tract, the experimental design was set up to use
pepsin and acidic condition for 30 min before applying a-amylase
in a dialysis system. Generally, pepsin cleaves peptide bonds of
proteins to form a range of shorter peptides and amino acids
(Brownlee, 2011), therefore the maltose and glucose levels detected
in a dialysis system were derived from a-amylase cleavage.
After digestion with a-amylase in a dialysis system for 15 min,
MNG-containing breads showed significantly reduced amounts of
maltose by 38% of control (Table 3) and decreased glucose by 37e
46% of control (Table 4). Maltose and glucose levels of 1 g/100 g
MNG-containing breads were slightly lower than that of 0.5 g/100 g
MNG-containing breads although there was no statistical signifi-
cance (p > 0.05). At the end of study period (at 180 min), MNG-
containing breads showed significantly lower amounts of maltose
(by 23e39%) and glucose (by 33e40%) than did control bread.
3.6. Scanning electron microscopy
The scanning electron microscopy of fresh control bread showed
two types of starch granules, spherical and oval shapes, protruding
from gluten matrix (Fig. 3). In contrast, fresh MNG-containing breads
showed smooth surface with starch granule adhesion in the gluten
matrix. Among MNG-containing breads, breads containing MNG

Table 4
Glucose contents in dialysate (mmol/L) of in vitro digestion in a bread e a-amylase-dietary fiber system.

Bread Glucose content in dialysate (mmol/L)

15 min 30 min 60 min 120 min 180 min

Control 3.6  0.15a 3.8  0.05a 4.1  0.11a 4.3  0.17a 4.6  0.13a
MNG extracted with 0.05 mol/L NaOH
0.5 g/100 g concentration 2.0  0.01bc 2.3  0.03bc 2.5  0.10bc 2.6  0.09bc 2.9  0.07bc
1.0 g/100 g concentration 1.9  0.01c 2.1  0.01c 2.3  0.03c 2.4  0.01c 2.7  0.03c
MNG extracted with 0.1 mol/L NaOH
0.5 g/100 g concentration 2.3  0.13b 2.4  0.06bc 2.7  0.12b 2.9  0.07b 3.0  0.10b
1.0 g/100 g concentration 2.2  0.05bc 2.3  0.01bc 2.5  0.15bc 2.7  0.18bc 2.9  0.03bc
MNG extracted with 0.2 mol/L NaOH
0.5 g/100 g concentration 2.3  0.07b 2.5  0.01b 2.8  0.09b 2.9  0.13b 3.0  0.10b
1.0 g/100 g concentration 2.2  0.15bc 2.3  0.17bc 2.5  0.10bc 2.6  0.11bc 2.8  0.13bc

Data present as mean  S.E. of three independent replicates.

aec
different letters following means within the same column indicate a significant difference at the p < 0.05 level.
 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494 491

Fig. 3. Scanning Electron Micrographs (1000) of control and MNG-containing breads. (A) control bread; (B) bread containing 0.5 g/100 g MNG extracted with 0.05 mol/L NaOH; (C)
bread containing 1 g/100 g MNG extracted with 0.05 mol/L NaOH; (D) bread containing 0.5 g/100 g MNG extracted with 0.1 mol/L NaOH; (E) bread containing 1 g/100 g MNG
extracted with 0.1 mol/L NaOH; (F) bread containing 0.5 g/100 g MNG extracted with 0.2 mol/L NaOH; (G) bread containing 1 g/100 g MNG extracted with 0.2 mol/L NaOH.

extracted with 0.05 mol/L NaOH had smooth surfactant with less
starch protruding from the matrix, with a less dense matrix than that
of bread containing MNG extracted with 0.1 and 0.2 mol/L NaOH.
After in vitro digestion for 3.5 h, control bread had much porous
appearance with less starch granule in the matrix indicating that a-
amylase had high ability to digest starch granules (Fig. 4). This
finding is consistent with the highest amount of maltose and
glucose after a-amylase digestion for 180 min in a dialysis system
(Tables 3 and 4). MNG-containing breads had less porosity and
more compact appearance with retention of undigested starch
granules than that of control. Specifically, 1 g/100 g MNG-
containing breads had undigested starch granules remained
intact with the matrix more than that of 0.5 g/100 g MNG-
containing bread after a-amylase digestion for 180 min (Fig. 4).
4. Discussion
Physical properties of hydrocolloid-containing breads depend
on the type and level of hydrocolloids added, as well as the level of
water incorporation. Skendi, Biliaderis, Papageorgiou, and
492 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494

Fig. 4. Scanning Electron Micrographs (1000) of control and MNG-containing breads digested with porcine pancreatic a-amylase for 180 min in a dialysis system. (A) control
bread; (B) bread containing 0.5 g/100 g MNG extracted with 0.05 mol/L NaOH; (C) bread containing 1 g/100 g MNG extracted with 0.05 mol/L NaOH; (D) bread containing 0.5 g/
100 g MNG extracted with 0.1 mol/L NaOH; (E) bread containing 1 g/100 g MNG extracted with 0.1 mol/L NaOH; (F) bread containing 0.5 g/100 g MNG extracted with 0.2 mol/L
NaOH; (G) bread containing 1 g/100 g MNG extracted with 0.2 mol/L NaOH.

Izydorczyk (2010) reported that addition of b-glucan in wheat flour
led to a decrease in loaf specific volume, in which the extent of
decrease depending on b-glucan level. Other reports studying the
effect of xanthan gum-containing bread showed that adding gum at
high concentration exhibited too high resistance and consistency
batter contributed to a limited gas cell expansion during proofing
(Lazaridou, Duta, Papageorgiou, Belc, & Biliaderis, 2007; Peressini,
Pin, & Sensidoni, 2011). In the present study, MNG-containing
breads had more open crumb cells led to significantly higher loaf
volume and loaf specific volume compared to that of control (Fig. 1
and Table 1). These results reflected that extracted MNGs enhanced
incorporation of gases during mixing or increased CO2 retention
during proofing. The results were in agreement with other reports
studying the effect of hydrocolloids on bread properties; an in-
crease in batter viscosity improves dough development and gas
retention, thereby increasing loaf volume (Sciarini, Ribotta, Leon, &
Perez, 2010).
Specific volume of both 0.5 g/100 g and 1 g/100 g MNG-
containing breads was not significantly different. These results
indicated that extracted MNGs at 1 g/100 g concentration might
 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494
bind a larger volume of water until breads was not fully expanded.
However, the specific volume of 1 g/100 g MNG-containing breads
was significantly higher than that of control. This indicated that
MNG did not tightly bind water until gluten was not properly hy-
drated. The results were opposite to other scientific studies
showing that b-glucan has high water affinity and limits the
available water within the paste mixture for the development of
the gluten network, which contributed to reduced height, reduced
loaf volume and increased crumb firmness (Cleary, Anderson, &
Brennan, 2007; Gaosong & Vasanthan, 2000; Gill et al., 2002;
Symons & Brennan, 2004). From the structure point of view, the
different characteristic of extracted MNGs may be beneficial for
application of food product development.
After 24 h of storage, the moisture content of 1 g/100 g MNG-
containing bread was significantly higher than that of 0.5 g/100 g
MNG-containing bread (Table 2). This difference, however, was not
found on day 2 and day 3 of storage. These results indicate that
increased concentration of MNG increased the ability of gum to
hold the free water but molecules were not tightly bound, therefore
excess water was lost in long term.
Generally, hydrophilic gums tightly bind the free water
contributing to increased water retention, therefore bread crumb
stays soft longer (Sharadanant & Khan, 2003). Changes in crumb
softness and moisture loss are factors to indicate staling process of
baked products during storage (Shittu, Aminu, & Abulude, 2009).
The ideal moisture level of baked product related to the softness is
between 35 and 40% (Shah, Shah, & Madamwar, 2006). The present
study showed that the rate of increased crumb firmness of MNG-
containing breads was significantly lower than that of control.
The moisture contents of MNG-containing breads during three-day
storage were higher than the ideal moisture level. Taken these re-
sults together, it indicates that the water binding capacity of
extracted MNGs could prevent water loss during bread storage. It
was proposed that viscous fiber has a larger water-holding capacity
than starch, thereby affecting decreased starch retrogradation and
consequently decreased crumb firmness (Purhagen, Sjoo, &
Eliasson, 2012). The results in this study were in agreement with
that of Shittu et al. (2009) who reported that moisture loss and
crumb firmness during bread storage were best reduced when 1 g/
100 g xanthan gum was added to bread formulation.
SEM of fresh control bread had starch granules that were pre-
dominant within protein matrix (Fig. 3). Addition of extracted
MNGs provided a continuous structure of starch granules
embedded into the protein matrix. The results were in agreement
with that of Brennan et al. (1996) who reported that gal-
actomannan dispersed and mixed intimately with the starch
granules and protein matrix to become a continuous structure
appearance.
The SEM of in vitro digestibility with a-amylase of MNG-
containing breads showed more undigested starch granules than
that of control (Fig. 4). In accordance with the results shown in
Tables 3 and 4, MNG-containing breads had significantly lower
levels of maltose and glucose than did control. Taken together,
these results suggest that extracted MNGs acted as a barrier to
prevent a-amylase to contact starch granules. Plasma glucose levels
rise about 10 min after a meal as a result of the absorption of car-
bohydrates ingested in the meal (Sudhir & Mohan, 2002). Accord-
ing to oral glucose tolerance test pattern, the highest peak of
plasma glucose levels after carbohydrate consumption is at 30e
60 min and gradually reduces to normal plasma level within
120 min. In order to determine the durable ability of extracted
MNGs to interfere a-amylase activity, bread e dietary fiber e a-
amylase in a dialysis system was continuously run for 180 min. This
study showed that the extracted MNGs in bread effectively pre-
vented starch digestion contributing to less change of glucose and
493
maltose levels in a dialysate during 180 min. In contrast, a control
bread had gradually increased levels of glucose and maltose up to
180 min. This finding is consistent with the previous report that
extracted MNGs interfered a-amylase activity in a starch solution e
a-amylase e dietary fiber system causing lower glucose content in
dialysate (Srichamroen & Chavasit, 2011b). The results of the pre-
sent study were in agreement with a study indicated that hydro-
colloid galactomannan may form a layer around starch granules,
which could then shield the starch granules from enzyme attack
(Slaughter et al., 2002). Brennan et al. (1996) also reported a lower
rate in the in vitro starch digestibility of guar galactomannan-
containing bread when incubated with porcine pancreatic a-
amylase compared to that of wheat bread.
5. Conclusion
Alkaline-extracted MNG added in bread formulation signifi-
cantly increased loaf volume and specific volume. This is correlated
with increased number of gas cells of the MNG-containing breads.
Addition of extracted MNGs in bread formulation significantly
reduced moisture loss and firmness of bread crumb after storage for
three days. The SEM of in vitro digestibility with a-amylase of MNG-
containing breads showed less porosity and more undigested
starch granules remained intact with the matrix. This finding was
consistent with the low level of glucose and maltose. Specifically,
MNG extracted with 0.05 mol/L NaOH had high potential to inter-
fere a-amylase activity in starch digestion process. The significance
of this study indicates that the durable ability of extracted MNGs to
interfere a-amylase activity in solid matrix was up to 180 min
regarding the time-experimental design.
Acknowledgments
The financial support, funding number R2553C194, from Nar-
esuan University Fund, Thailand is gratefully acknowledged. The
author would like to thank Mrs. Prakaytip Kot-asa, Faculty of Sci-
ence, Naresuan University for her SEM technical support.
References
Abu-Ghoush, M., Herald, T. J., Dowell, F., Xie, F., Aramouni, F. M., & Walker, C. (2008).
Effect of antimicrobial agents and dough conditioners on the shelf-life exten-
sion and quality of flat bread, as determined by near-infrared spectroscopy.
International Journal of Food Science and Technology, 43, 365e372.
American Association of Cereal Chemists (AACC). (2000). Approved methods of the
AACC (10th ed.). St Paul, MN: The Association (Method 44e15A).
Anderson, J. W., Akanji, A. O., & Randles, K. M. (2001). Treatment of diabetes with
high-fibre diets. In G. A. Spiller (Ed.), CRC handbook of dietary fibre in human
nutrition (3rd ed.) (pp. 373e400). New York: CRC Press.
Blackburn, N. A., Redfern, J. S., & Jarjis, H. (1984). The mechanism of action of guar
gum in improving glucose tolerance in man. Clinical Science, 66(3), 329e336.
London.
Brennan, C. S., Blake, D. E., Ellis, P. R., & Schofield, J. D. (1996). Effects of guar gal-
actomannan on wheat bread microstructure and on the in vitro and in vivo
digestibility of starch in bread. Journal of Cereal Science, 24, 151e160.
Brownlee, I. A. (2011). The physiological roles of dietary fibre. Food Hydrocolloids, 25,
238e250.
Ceriello, A. (2005). Postprandial hyperglycaemia and diabetes complications: is it
time to treat? Diabetes, 54, 1e7.
Cleary, L. J., Anderson, R., & Brennan, C. S. (2007). The behaviour and susceptibility
to degradation of high and low molecular weight barley b-glucan in wheat
bread during baking and in vitro digestion. Food Chemistry, 102, 889e897.
Czuchajowska, Z., Pomeranz, Y., & Jeffers, H. C. (1989). Water activity and moisture
content of dough and bread. Cereal Chemistry, 66, 128e132.
Gaosong, I., & Vasanthan, T. (2000). Effect of extrusion cooking on the primary
structure and water solubility of b-glucan from regular and waxy barley. Cereal
Chemistry, 77, 396e400.
Gill, S., Vasanthan, T., Ooraikul, B., & Rossnagel, B. (2002). Wheat bread quality as
influenced by the substitution of waxy and regular barley flours in their native
and extruded forms. Journal of Cereal Science, 36, 219e237.
Heine, R. J., & Dekker, J. M. (2002). Beyond postprandial hyperglycaemia: metabolic
factors associated with cardiovascular disease. Diabetologia, 45, 461e475.
494 A. Srichamroen / LWT - Food Science and Technology 59 (2014) 486e494
Huttner, E. K., & Arendt, E. K. (2010). Recent advances in gluten-free baking and the
current status of oats: a review. Trends in Food Science & Technology, 21, 303e
312.
Lazaridou, A., Duta, D., Papageorgiou, M., Belc, N., & Biliaderis, C. G. (2007). Effects of
hydrocolloids on dough rheology and bread quality parameters in gluten-free
formulation. Journal of Food Engineering, 79, 1033e1047.
Peressini, D., Pin, M., & Sensidoni, A. (2011). Rheology and breadmaking perfor-
mance of rice-buckwheat batters supplemented with hydrocolloids. Food Hy-
drocolloids, 25, 340e349.
Purhagen, J. K., Sjoo, M. E., & Eliasson, A. C. (2012). Fibre-rich additives e the effect
on staling and their function in free-standing and pan-baked bread. Journal of
the Science of Food and Agriculture, 92, 1201e1213.
Rainbird, A. L., Low, A. G., & Zebrowska, T. (1984). Effect of guar gum on glucose and
water absorption from isolated loops of jejunum in conscious growing pigs.
British Journal of Nutrition, 52(3), 489e498.
Rosell, C. M., Rojas, J. A., & Benedito de Barber, C. (2001). Influence of hydrocolloids
on dough rheology and bread quality. Food Hydrocolloids, 15, 75e81.
Sabanis, D., & Tzia, C. (2011). Selected structural characteristics of HPMC-containing
gluten free bread: a response surface methodology study for optimizing quality.
International Journal of Food Properties, 14, 417e431.
Sciarini, L. S., Ribotta, P. D., Leon, A. E., & Perez, G. T. (2010). Influence of gluten free
flours and their mixtures on batter properties and bread quality. Food and
Bioprocess Technology, 3, 577e585.
Shah, A. R., Shah, R. K., & Madamwar, D. (2006). Improvement of the quality of
whole wheat bread by supplementation of xylanase from Aspergillus foetidus.
Bioresource Technology, 97, 2047e2053.
Sharadanant, R., & Khan, K. (2003). Effect of hydrophilic gums on the quality of
frozen dough: II bread characteristics. Cereal Chemistry, 80(6), 773e780.
Shittu, T. A., Aminu, R. A., & Abulude, E. O. (2009). Functional effects of xanthan
gums on composite cassava wheat dough and bread. Food Hydrocolloids, 23(8),
2254e2260.
Skendi, A., Biliaderis, C. G., Papageorgiou, M., & Izydorczyk, M. S. (2010). Effects of
two barley b-glucan isolates on wheat flour dough and bread properties. Food
Chemistry, 119(3), 1159e1167.
Slaughter, S. L., Ellis, P. R., Jackson, E. C., & Butterworth, P. J. (2002). The effect of guar
galactomannan and water availability during hydrothermal processing on the
hydrolysis of starch catalysed by pancreatic a-amylase. Biochimica et Biophysica
Acta, 1571, 55e63.
Srichamroen, A., & Chavasit, V. (2011a). Rheological properties of extracted malva
nut gum (Scaphium scaphigerum) in different conditions of solvent. Food Hy-
drocolloids, 25(3), 444e450.
Srichamroen, A., & Chavasit, V. (2011b). In vitro retardation of glucose diffusion with
gum extracted from malva nut seeds produced in Thailand. Food Chemistry, 127,
455e460.
Sudhir, R., & Mohan, V. (2002). Postprandial hyperglycaemia in patients with type 2
diabetes mellitus. Treatments in Endocrinology, 1(2), 106e116.
Symons, L. J., & Brennan, C. S. (2004). The influence of (1-3) (1-4)-b -D glucan-rich
fractions from barley on the physicochemical properties and in vitro reducing
sugar release of white wheat breads. Journal of Food Science, 69(6), C463eC467.
Torsdottir, I., Alpsten, M., Anderson, H., & Einansson, S. (1989). Dietary guar gum
effects on postprandial blood glucose, insulin and hydroxyproline in humans.
Journal of Nutrition, 119, 1925e1931.
Woolnough, J. W., Monro, J. A., Brennan, C. S., & Bird, A. R. (2008). Simulating human
carbohydrate digestion in vitro: a review of methods and the need for stan-
dardization. International Journal of Food Science and Technology, 43, 2245e
2256.