Food Hydrocolloids 57 (2016) 209e216

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Comparative studies of konjac flours extracted from Amorphophallus

guripingensis and Amorphophallus rivirei: Based on chemical analysis
and rheology
Qing Huang a, Weiping Jin a, Shuxin Ye a, Ying Hu a, Yuntao Wang a, Wei Xu a, Jing Li a,
Bin Li a, b, c, *
a
College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
b
Hubei Collaborative Innovation Centre for Industrial Fermentation, Hubei University of Technology, Wuhan 430068, China
c
Key Laboratory of Environment Correlative Dietology (Huazhong Agricultural University), Ministry of Education, China

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

Article history: Konjac glucomannan (KGM) has been widely used in food industry, which is commonly extracted from
Received 10 May 2015 Amorphophallus rivirei. In this paper, Amorphophallus guripingensis was found to be another good
Received in revised form resource of extracted KGM. The comparative studies, based on chemical composition analysis and
15 January 2016
rheological measurements, between those two different resources were investigated. Results showed
Accepted 20 January 2016
Available online 21 January 2016
that refined A. guripingensis flour (AGF) contained 61.35% KGM, which was lower than that of refined ARF
(72.80%). But, the viscosity-increased speed of AGF was faster than ARF. To be specific, even the viscosity
of 3.5 wt% AGF solution was half of ARF, while they had similar viscosity values at a concentration of
Keywords:
Amorphophallus rivirei
4.5 wt%. Besides, viscosity change of the refined AGF with pH was more pronounced under strong acid
Amorphophallus guripingensis condition. Meanwhile, thermo-gelling temperature of 4.0 wt% AGF solution was 85
C, lower than that of
Refined flour ARF. These results demonstrated that AGF solutions were sensitive to processing factors, such as pH and
pH-sensitivity temperature as compared to A. rivirei. Those properties endows AGF more valuable applications in food
Rheological properties industry and it has great potential to be a pH-sensitive matrix.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction
Refined konjac flour (refined KF), a kind of powder that is pu-
rified from the tuber of amorphophallus, is mainly farmed in
Southeast Asia and India (Li, Xia, Wang, & Xie, 2005). After har-
vesting, fresh konjac tuber is washed, sliced, dried and grounded.
Then the crude powder is further purified by mechanical ways or/
and alcohol wash so as to remove impurities and thus the refined
konjac flour was obtained (Liu, 2004, chap. 19). Refined KF con-
tained a large amount of konjac glucomannan (KGM), a natural
neutral polysaccharide consisting of mannose and glucose with b-
1-4 linkage. Its molar ratio as well as acetyl groups along the KGM
backbone might differ depending on the original source of KGM
(An, Thien, Dong, Dung, & Du, 2010; Chen, Zong, & Li, 2006;
Nguyen, Do, Nguyen, Pham, & Nguyen, 2011).
* Corresponding author. College of Food Science and Technology, Huazhong
Agricultural University, Wuhan 430070, China.
E-mail address: libinfood@mail.hzau.edu.cn (B. Li).
China is the first country to study and utilize konjac (Chua,
Baldwin, Hocking, & Chan, 2010). In the past, konjac flour has
been consumed as medicinal material and traditional food in the
form of konjac jelly, tofu, noodles (Joyce Keithley and Swanson,
2005). After the methodology for purifying konjac was proposed
by Mashiko, the application of refined KF was greatly broadened
(Chua et al., 2010). The refined konjac flour occupies the majority of
the konjac market. Every year, about 1,200,000 tons refined konjac
flour is produced in the world (Sheng & Teng, 2008). Due to its high
water-absorbing capacity, konjac flour is commonly used as thick-
ening and gelling agent (Saha & Bhattacharya, 2010). Recently,
much more attention has been paid to refined KF as functional
products for its potential value, being a dietary fiber with functions
of lowering blood sugar, removing blood fat and cholesterol, as well
as regulating intestinal flora (Jime
nez-Colmenero et al., 2012; Joyce
Keithley and Swanson, 2005; Li et al., 2005; Sood, Baker, &
Coleman, 2008). Moreover, application in the field of material and
medicine also achieved some degree of development, such as ex-
cipients for floating system, films for wound dressing and

http://dx.doi.org/10.1016/j.foodhyd.2016.01.017
0268-005X/© 2016 Elsevier Ltd. All rights reserved.
210 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216
microspheres for low-cost desalting (Huang, Chu, Huang, Wu, &
Tsai, 2015; Ji & Deng, 2011; Xiong et al., 2014). These recent de-
velopments in food, material and medical fields give rise to the
increasing demand for konjac and it is urgent to fully characterize
the properties of konjac in order to fulfill its market potential.
Konjac is an Araceae perennial herb, and there are about 170
kinds of konjac in the globe as previously reported (Hetterscheid &
Ittenbach, 1996). But konjac flours extracted form Amorphophallus
rivirei and A. albus dominated the konjac market in the world. In
order to satisfy the growing demand for konjac flour, exploring new
varieties, such as Amorphophallus guripingensis, became an inevi-
table trend. Compared with the commercial varieties (A. rivirei and
A. albus), A. guripingensis has several outstanding advantages, such
as high propagation coefficient, disease resistance and adaptability
to hot or humid environment, which qualify it a potential com-
modity. But the rarely research about A. guripingensis seriously
limited the application and development of new food ingredients.
Although differences in chemical composition and KGM structure
between species have been reported previously (An et al., 2010;
Fang & Wu, 2004), rheological properties between varieties was
scarcely conducted and they directly influenced practical usage of
konjac flour in the source of A. guripingensis.
The present study was aimed to understand the characteristics
of A. guripingensis refined flour, enrich the basic knowledge of its
rheological properties and broaden the application. In the
comparative studies, chemical compositions and rheology are two
major aspects for investigation. This study would provide theo-
retical and practical information on developing new konjac based
food ingredients.
2. Materials and methods
2.1. Materials
Two kinds of refined konjac flour that extracted from
A. guripingensis and A. rivirei were supplied by QiangSen Biotech
Co (Wuhan, China). In chemical analysis and rheological testes,
they were used directly without any further purification. In the
KGM molecular structure measurements, KGM was purified from
two kinds of refined konjac flour until the purity reached at least
90%. All of other chemicals used were of analytical grade and
purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai,
China).
2.2. Chemical compositions analysis
For the two kinds of refined KF, the contents of moisture, ash,
protein, and soluble sugar were analyzed according to AOAC
methods (Horwitz, 2000). The dry basis content of KGM was
measured as previously reported (Xu et al., 2014).
2.3. Molecular parameters of konjac glucomannan
Preparation and purification of KGM were performed accord-
ing to (Jian et al., 2013). Briefly, 10 g refined KF was washed with
50 ml 50% ethanol containing 0.1% sodium azide for three times
so that water-soluble impurities were removed. Then the flour
was dried in air. After that, it was placed in 50 ml mixed solvent
of absolute aether/absolute ethanol [2:1(V/V)] at 40
C with
mechanical stirring for 8 h in order to remove grease. Secondly,
the degreased sample was dissolved in distilled water to be a 0.6%
(W/V) solution with mechanical stirring at 30
C water bath for
2 h. The fibrin and other insoluble compositions were separated
by over-speed freezing centrifuge at a rate of 16,000 r/min for
20 min. Then the supernatant was collected and appropriate
amount of amylase was added at room temperature till enzymatic
hydrolysis was finished for removing amylum. Besides, protein
was removed by the enzymatic hydrolysis, combining Sevag
method with centrifugation for five times, and then water phase
was collected. After that, the sample was deposited by adding
same volume 95% ethanol (V/V) and then washed with absolute
ethanol and aether. Finally, KGM was obtained after vacuum
drying.
The molecular weight of KGM was determined by the method of
(Jin et al., 2014). In brief, 0.06 wt% purified KGM was prepared by
mechanical stirring and diluted into a series of concentration with
0.2 M NaCl solution. Before determination, the sample was filtered
through a 0.45 mm Millipore filter to remove insoluble impurities.
Then viscosity-average molecular weight of purified KGM was
measured using an Ubbelohde viscometer (Liangjing Co., Shanghai,
China) at 25 ± 0.1
C. All the measurements were performed in six
replicates.
The acetyl content was quantified by the modified means of (T.
Zhang, Xue, Li, Wang, & Xue, 2015). Four grams dried purified KGM
was placed in a 150 ml Erlenmeyer flask with a stopper and 40 ml
75% ethanol was added. The flask was shaken in a thermostatic
oscillator (its agitation speed was 150 rpm) for 30 min at 40
C.
After that, 5 ml 0.5 M NaOH was added and the flask was shaken
continually at 40
C for 24 h. The excess alkali was titrated back
with 0.1 M HCl using phenolphthalein as an indicator. The blank, no
KGM sample added, was titrated in parallel. All experiments were
carried out three times at least.
2.4. Rheological properties
2.4.1. Samples preparation
To investigate the effect of concentration on rheological
behavior, two kinds of refined KF of various concentrations
including 3.5 wt%, 4.0 wt%, 4.5 wt%, 5.0 wt%, 5.5 wt% (w/w)
were completely dissolved in distilled water with mechanical
stirring at 30
C for 2 h. Then the solution was filtered with
300 mesh gauze to remove insoluble impurities, and degassed by
vacuum.
To study the effect of pH on rheological behavior, the concen-
tration was fixed at 4.0 wt%. The solution was prepared with similar
methods stated before. Different pH values including 2, 4, 6, 8, 10
were adjusted with 0.1 M sodium hydroxide and 0.1 M hydrochloric
acid.
2.4.2. Rheological measurements
All rheological measurements were performed using TA AR
2000ex controlled strain rheometer with peltier plate system.
Freshly prepared samples were equilibrated at 25
C for 20 min
before rheological measurements. Steady-state flow curves were
obtained using 2
cone plates (40 mm diameter and 0.052 mm gap)
at 25
C, over the shear rate range of 0.1e300 s1.
Dynamic oscillatory measurements were performed using par-
allel plates (40 mm diameter, 1 mm gap). Frequency sweeps were
conducted in the 0.1e500 rad/s range and the strain was fixed at 2%
in order to assure the working conditions within the linear visco-
elastic region. Temperature sweeps were carried out from 25
C to
90
C at a constant heating rate of 2
C/min. Meanwhile, the strain
and frequency were fixed at 2% and 6.283 rad/s respectively. After
the sample was placed into the measuring device, a thin layer of
paraffin oil was spread over the outer edge of the sample to prevent
moisture evaporation. The measurements were conducted in trip-
licate. Viscosity, storage modulus (G0 ) and loss modulus (G00 ) were
recorded.
 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216
3. Results and discussion
3.1. Composition analysis
The physicochemical properties were significantly affected by
compositions of refined KF. Their main compositions were pre-
sented in Table 1, including the content of KGM, moisture, ash,
protein and soluble sugar. It was suggested that the content of KGM
in refined AGF (61.347 ± 2.293%) was lower than that of ARF
(72.801 ± 1.945%). And the other content (not determined), espe-
cially insoluble polysaccharides, such as starch, cellulose (the main
component of the cell wall), in refined AGF was higher compared
with refined ARF. Besides, the parameters of konjac glucomannan
from two species were shown in Table 2. It was observed that the
molecular weight of KGM in refined AGF and ARF were 1.236  106
and 1.669  106, respectively. Furthermore, the molecular weight
was directly associated with solution viscosity (Jin et al., 2014) and
various viscosities were needed in different foods so that the above
data might contribute to guide the industrial usage of konjac flour.
What's more, refined AGF possessed lower acetyl groups compared
with ARF and acetyl content was closely related to gel-forming
property (Du, Li, Chen, & Li, 2012; Gao & Nishinari, 2004a,
2004b). It was commonly considered that KGM with lower acetyl
groups was easier to form a gel. These differences in composition
and parameters of KGM were attributed to species, regions, soil
environment and so on (Liu, 2004, chap. 19).
3.2. Rheological analysis
3.2.1. Effect of concentration on rheological properties
The flow curves of the refined AGF and ARF at the different
concentrations were studied and the results were presented in
Fig. 1. The sample showed a remarked shear-thinning behavior at
low shear rate and the viscosity finally remained constant at high
shear rate, a typical pseudo-plastic fluid. Similar rheological
observation had also been reported in previous study about puri-
fied KGM (Wang et al., 2012). This type of behavior was expected for
most polymer solutions (Hua, Wang, Yang, Kang, & Zhang, 2015;
Wu, Ding, Jia, & He, 2015). The phenomenon could be explained
by that shear force disturbed the polysaccharides' entangled
molecule networks and decreased the effective size of polymers
(Bhandari, Singhal, & Kale, 2002). But molecular chains were
rearranged and aligned with the orientation of the shear force,
resulting in the unchanged viscosity (Hua et al., 2015).
The steady shear viscosity of the two kinds of refined KF as a
function of shear rate for different concentrations varying from
3.5 wt% to 5.5 wt% was presented in Fig. 1a and b. It was demon-
strated that the shear-thinning behavior was more remarkable at
25
C with the increase of concentration. This phenomenon was in
good agreement with those found for other solutions (Gong et al.,
2012; Khounvilay & Sittikijyothin, 2012; Torres, Hallmark, &
Table 1
Main compositions of refined konjac flour (%).
A. guripingensis A. rivirei
a
KGM (%) 61.35 ± 2.293 72.80
Moisture (%) 11.69 ± 0.009 11.97
Ash (%) 4.262 ± 0.002 4.765
Protein (%) 1.458 ± 0.006 4.685
Soluble sugar (%) 1.878 ± 0.015 3.310
Othersb e e
e Not determined.
a
Dry basis content.
b
Starch, cellulose (the main component of the cell wall) and so on.
211
Table 2
Main parameters of konjac glucomannan.
A. guripingensis A. rivirei
Molecular weight of KGM 1.236  106 1.669  106
Acetyl content (%) 1.342 ± 0.033 2.285 ± 0.026
Wilson, 2014). Moreover, the viscosity of the refined KF solution
increased obviously with increase of concentration. This phenom-
enon was mainly attributed to intermolecular interactions between
konjac glucomannan (the main component of refined KF), such as
hydrogen bonds, caused by massive hydroxyl groups. With
increasing concentration, the molecular chains within the unit
space increased, resulting in obvious stronger intermolecular
hydrogen bonds and then more entangled points between molec-
ular chains appeared. Finally, the more entangled three-
dimensional network formed, which led to the higher viscosity
(Hua et al., 2015; Wu et al., 2015). When the concentration was
lower than 4.5 wt%, the shear viscosity of refined AGF was lower
than that of refined ARF, while at 4.5 wt% (or above) concentration,
the shear viscosity of both KF were equivalent, which might be due
to the difference between the plant cell wall of two species. At low
concentration, the friction force between the A. guripingensis par-
ticles cell was so small that some of the particles did not swell and
extend completely. Thus small parts of undissolved KGM molecules
were also removed by filtration along with impurities together.
However, A. rivirei particles were in the opposite situation.
Accordingly, the varieties exhibited differences on the viscosity.
The frequency dependence of AGF and ARF solutions' G0 and G00
with various concentrations from 3.5 wt% to 5.5 wt% at 25
C was
presented in Fig. 2. It was found that both of G0 and G00 rose with
increasing frequency and a crossover could be detected, which
could be explained by assuming that the entanglement of molec-
ular chain was transient instead of permanent. At low frequencies,
KGM molecular chains disentangled during a long period oscilla-
tion, so that G00 was higher than G0 , indicating a liquid-like behavior.
Whereas at higher frequencies, molecular chains had no enough
time to disentangle during a short period of oscillation. Therefore,
G0 was higher than G00 , showing a solid-like behavior (Du et al.,
2012; Jin et al., 2014).
In Fig. 2, with an increase of concentration, the crossover points
of the refined AGF and ARF shifted to lower frequency from
20.508 rad/s to 5.290 rad/s and from 2.746 rad/s to 1.721 rad/s,
respectively, which indicated the enhanced entangled network (Jin
et al., 2014). Similar phenomenon was observed for other hydro-
colloids, such as tamarind seed gum (Khounvilay and Sittikijyothin,
2012) and Tara gum (Wu et al., 2015). It was contributed to that
molecular chains of KGM got closer to each other at higher con-
centration and that the probability of the formation of a junction
zone was higher (Cai & Zhang, 2006). Moreover, the crossover
points of refined ARF (Fig. 2a) were lower than AGFs' (Fig. 2b) at the
same concentration. The larger molecular weight and higher con-
tent of KGM in the refined ARF compared with AGF might account
for these phenomena (Cai and Zhang, 2006; Jin et al., 2014; Zhong
et al., 2015). As the number of KGM molecules and length of the
KGM chain increased, their relaxation capacity decreased, resulting
in easier entanglement.
± 1.945 Temperature sweeps were used to explain behavior upon
± 0.034
± 0.008
heating in tested system. The effect of temperature on G0 and G00 of
± 0.013 the refined AGF and refined ARF at various concentrations upon
± 0.041 heating from 25
C to 90
C was shown in Fig. 3a and Fig. 3b,
respectively. In Fig. 3a, with increasing temperature, G0 and G00
decreased and departed. Meanwhile, G0 was lower than G00 ,
showing a fluid-like behavior and it was more pronounced at
212 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216

Fig. 1. Viscosity-shear rate profiles of different concentrations from two species at shear rate from 0.1 s1 to 300 s1 (T ¼ 25
C). (a) the refined AGF; (b) the refined ARF.

Fig. 2. Frequency dependence of G0 and G00 for the refined KF solution at various concentrations from 3.5 wt% to 5.5 wt%. (a) the refined AGF; (b) the refined ARF. The data were
shifted along the vertical axis by 10a to avoid overlapping.

Fig. 3. Temperature dependence of G0 and G00 for the refined KF solution at various concentrations from 3.5 wt% to 5.5 wt%. (a) the refined AGF; (b) the refined ARF. The data were
shifted along the vertical axis by 10a to avoid overlapping.

higher temperature. It might be supported by the weakening of the
hydrogen bond, regarded as primary molecular force (T. Zhang
et al., 2015) and the declining of chain entanglements caused by
thermal disruption (Arancibia et al., 2015). As heating continued, G0
got closer to G00 and finally became higher than G00 when the con-
centration was above 4.5 wt%. It was demonstrated that the
behavior of the refined AGF solution was shifted from fluid-like to
solid-like. In addition, fluids-like behavior was maintained over the
whole tested temperature interval (25e90
C) at low concentra-
tions, while the crossover points (G0 ¼ G00 ) were detected at con-
centration higher than 4.5 wt% at particular temperatures (86.2
C,
84.7
C, 79.9
C, respectively). And there was a tendency to decrease
in temperature (crossover happened) as concentration increased. It
might be due to the low acetyl content and strong hydrophobic
interactions (predominated molecular force at an increasing tem-
perature), giving rise to the aggregation of molecular chains of KGM
and the subsequent formation of junction zones or molecular chain
clusters (Du et al., 2012; T. Zhang et al., 2015). Nevertheless, the
temperature-dependent G0 and G00 of the refined ARF were different
(Fig. 3b). Both G0 and G00 decreased with increase of temperature
from 25
C to 90
C. At low temperature, G0 was higher than G00 at all
investigated concentrations, showing a solid-like behavior, since a
weak gel network was already formed by hydrogen bonds. How-
ever, when the temperature rose unceasingly, hydrogen bonds
were destroyed, resulting in the disruption of network and the
weak gel melt to sol (G0 < G00 ).
 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216
3.2.2. Effect of pH on rheological properties
KGM was a neutral polysaccharide so that the rheological
properties of the sample should not be affected by pH among the
tested range (2e10) obviously. But the results of the refined KF
were different. Viscosity-shear rate profiles of the refined KF were
presented in Fig. 4. It was worth noting that the samples still kept a
shear-thinning behavior at various pH values as a result of the
breakup of intermolecular entanglements. This phenomenon was
similar to the effect of concentration. In Fig. 4a, the viscosity of the
refined AGF was highly dependent on pH. However, there was
almost no effect of pH on viscosity of refined ARF except for strong-
acid condition (Fig. 4b). As for the refined AGF, the viscosity un-
derwent a sharp increase from 348.8 Pa.s to 1522.0 Pa.s when pH
decreased from 4 to 2 and increased obviously from 69.7 Pa.s to
594.5 Pa.s with pH increasing from 8 to 10. Besides, the viscosity
changed markedly at pH among 4 to 8. Since there was still a small
portion of other ingredients and the extent affected by the external
environment were also different. At acidic condition, lowing pH
could cause the protonation of carboxyl groups on polymers, just
like protein in the refined KF, facilitating the formation of inter-
molecular hydrogen bonding and leading to the increase of vis-
cosity (Hua et al., 2015). On the other hand, KGM molecular chains
were broken by strong acid, resulting in the decrease of viscosity
(Kato & Matsuda, 1969). At alkaline condition, the hydroxyl groups
on the polymer chains were ionized (Wu et al., 2015), giving rise to
the disruption of intra and intermolecular hydrogen bonding, thus
viscosity of sample solution decreased. For another, deacetylation
occurred (Du et al., 2012; T. Zhang et al., 2015), resulting in the self-
association of KGM chains and giving rise to the higher viscosity.
Consequently, the difference of viscosity at various pH values might
be the combined results of above described factors. Nevertheless,
the exact reasons for these behaviors were not clear so far and
much more studies were needed.
G0 and G00 of two refined KF solutions at different pH as a
function of frequency were shown in Fig. 5. For all refined KF so-
lutions, G0 and G00 increased with an increase of frequency. More-
over, frequency at the intersection of refined AGF was larger than
that of ARF at same pH value, except under pH 2. Compared with
refined ARF, lower KGM content and smaller molecular weight in
AGF (Tables 1 and 2tbl2) might explain this phenomenon. In Fig. 5a,
the crossover point of G0 and G00 shifted to lower frequency at low
pH (0.571 rad/s at pH 2), owing to the enhanced intermolecular
hydrogen bonds, intra and intermolecular entanglements and as-
sociations (Bu, Kjøniksen, & Nystro € m, 2005). Nevertheless, the
crossover point changed slightly at high pH values. This might be
attributed to the reason that electrostatic interactions and
hydrogen bonding interactions changed with the media pH values.
Fig. 4. Viscosity-shear rate profiles of two kinds of refined KF solution (4.0 wt%) under different pH at shear rate from 0.1 s1 to 300 s1 (T ¼ 25
C). (a) the refined AGF; (b) the
refined ARF.
213
Most importantly, the magnitude of overall forces was slightly
changed. Whereas the intersection value of each frequency sweep
for refined ARF was almost unchanged (Fig. 5b), probably because
pH-insensitive KGM possessed the vast majority of the refined ARF.
The thermo-rheological behaviors of the refined KF at different
pH were explored (Fig. 6). It was suggested that the influence of pH
on rheological behavior of refined AGF was more remarkable than
that of ARF. As it was clearly shown, in Fig. 6a, during the initial
heating process, G0 and G00 at pH varying from 4 to 10 decreased
with increasing temperature and G0 was lower than G00 , indicating a
viscous behavior. However, as the temperature rose continually, G0
and G00 increased and the crossover point was observed, showing an
elastic behavior at high temperature. Therefore, there was a solegel
transition in the whole range of investigated temperature. This
phenomenon could be explained by the nature of thermal-induced
gelation of konjac glucomannan (H. Zhang et al., 2001). Moreover,
the crossover point was pH-dependent, owing to the different level
of entanglements and associations at pH ranging from 4 to 10.
However, when pH value was low to 2, the weak gel was formed
already and the gelesol transition happened until the temperature
rose to 45.9
C (Gigli, Garnier, & Piazza, 2009; Sittikijyothin,
Sampaio, & Gonçalves, 2007). Under strong acid condition, elec-
trostatic repulsion was suppressed and hydrogen-bonding in-
teractions dominated, which promoted the formation of a three-
dimensional network structure, i.e. a weak gel was formed. When
the temperature rose to 45.9
C, hydrogen-bonding interactions
were disrupted, the network structure was destroyed and the weak
gel was transformed into sol.
As for refined ARF (Fig. 6b), when the temperature rose gradu-
ally within the tested temperature (from 25
C to 90
C), G0 and G00
of the sample at every pH decreased and an intersection was
detected at about 50
C and gelesol transition happened, which
was similar to the refined AGF at pH 2. Moreover, the temperature
of the crossover detected was higher than that of refined AGF at pH
2. This might due to the higher KGM content and larger molecular
weight in refined ARF, thus entanglements and associations
occurred even at weak acidic or alkaline conditions. Much more
ordered network was formed and therefore more energy to destroy
this network structure was needed.
In fact, it was universally acknowledged that the addition of
alkali results in the deacetylation of KGM and finally leads to
gelation at elevated temperature, which was a time-independent
process. However, during this process, konjac concentration, mo-
lecular weight, degree of acetylation, alkali concentration, tem-
perature and heating time played an important role (Gao &
Nishinari, 2004a, 2004b; Hirai, 1954; Huang, Takahashi,
Kobayashi, Kawase, & Nishinari, 2002; Zhang et al., 2001). These
214 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216
Fig. 5. Frequency dependence of G0 and G00 of two kinds of refined KF solution (4.0 wt%) at different pH. (a) the refined AGF; (b) the refined ARF. The data were shifted along the
vertical axis by 10a to avoid overlapping.
Fig. 6. Temperature dependence of G0 and G00 for two kinds refined KF solution (4.0 wt%) under different pH from 2 to 10. (a) the refined AGF; (b) the refined ARF. The data were
shifted along the vertical axis by 10a to avoid overlapping.
factors finally determined the gelation time. Besides, there are
distinct differences both in the main composition of the refined
konjac flour and the main parameters of konjac glucomannan
extracted from two species (Tables 1 and 2tbl2). Hence, the gelation
behaviors might vary from species to species. In Fig. 6a, the gels of
refined AGF (pH ¼ 4e10) were formed primarily by hydrophobic
interactions at elevated temperature within the short time, which
were thermo-irreversible gels (“real gel”). Furthermore, due to its
relative low acetyl content, hydrophobic interaction was strength-
ened (Du et al., 2012), thus the gels were also formed in the tested
time even at weak acidic condition without completely deacetyla-
tion. As for the refined ARF (in Fig. 6b) and refined AGF (pH ¼ 2), the
gels were formed by hydrogen bonding interactions at low tem-
perature, which were thermo-reversible gels (“fake gel”). Once the
gel was subject to the high temperature, the weak gel was dis-
rupted. But with the increasing temperature within the tested
range, the thermo-irreversible gels were not formed within the
short time for the sake of high acetyl content.
3.2.3. Effect of temperature on rheological properties
To study the effect of temperature on rheological behavior of
refined KF, steady shear viscosity was measured at different tem-
peratures and the results were presented in Fig. 7. For the 4.0 wt%
refined KF solution (Fig. 7a and b), a typical pseudo-plastic behavior
was also observed within the investigated temperatures at shear
rate ranging from 0.1 s1 to 300 s1. Besides, it was obvious that
when the temperature was below 65
C, the viscosity of refined AGF
were less than that of ARF and both of them decreased with
increasing temperature from 1692.0 Pa.s to 239.6 Pa.s and
968.9 Pa.s to 197.1 Pa.s, respectively. These declination in viscosity
could be explained by the increased molecular distance (Kim & Yoo,
2009; Ma, Lin, Chen, Zhao, & Zhang, 2014; Xiao, Tong, & Lim, 2012)
and reduced hydrogen bonding force (Du et al., 2012) as a result of
elevated temperature. As for hydrophilic polymer solutions, it was a
common phenomenon that viscosity dropped with raising tem-
perature, such as carboxymethyl guar gum (Gong et al., 2012;
Shafiei-Sabet, Hamad, & Hatzikiriakos, 2012). When the tempera-
ture reached up to 85
C, the viscosity of ARF also decreased
continually. Conversely, viscosity of AGF increased sharply, reach-
ing 477.8 Pa.s, and even exceeded that of ARF. This might due to the
relative low content of acetyl groups along the backbone of KGM in
refined AGF so that the hydrophobic interaction between molecules
enhanced significantly at 85
C, resulting in the increased viscosity.
The dynamical oscillatory frequency sweep tests were carried
out to determine the viscoelastic behavior at angular frequencies
ranging from 0.1 to 500 rad/s. G0 and G00 for 4.0 wt% refined KF at
different temperatures were found to increase with increasing
angular frequency (Fig. 8). Furthermore, in Fig. 8a, at temperature
below 85
C, G00 was larger than G0 at low frequencies, showing the
fluid-like behavior. There was a tendency for G0 and G00 to approach
each other and the crossover was detected at high frequency. When
the frequency increased continually, G00 was lower than G0 , indi-
cating that the solid-like behavior dominated in the tested system
(Du et al., 2012; Jin et al., 2014). Interestingly, G0 and G00 increased
rapidly and showed a significant frequency-independent plateau at
85
C. Besides, G0 was greater than G00 , showing an elastic behavior
at all tested frequencies. This implied that the stable gel network
structure was formed (Cai and Zhang, 2006). In Fig. 8b, the cross-
over frequency of G0 and G00 shifted gradually to a lower value with
an increase of temperature, from 1.184 to 17.241 rad/s, indicating an
 Q. Huang et al. / Food Hydrocolloids 57 (2016) 209e216
Fig. 7. Viscosity-shear rate profiles of two kinds of refined KF solutions (4.0 wt%) at different temperature with shear rate ranging from 0.1 s1 to 300 s1. (a) the refined AGF; (b) the
refined ARF.
Fig. 8. Frequency dependence of G0 and G00 of two kinds of refined KF solution (4.0 wt%) at different temperatures. (a) the refined AGF; (b) the refined ARF. The data were shifted
along the vertical axis by 10a to avoid overlapping.
enhancement of temporary network structure of polymer chains
(Du et al., 2012).
4. Conclusions
The refined AGF (61.35 ± 2.293%) possessed lower content of
KGM in comparison with that of refined ARF (72.80 ± 1.945%). The
content of acetyl groups along the backbone of KGM in AGF
(1.342 ± 0.033%) was also lower than that of ARF (2.285 ± 0.026%).
But the molecular weights of KGM from two species were similar,
1.236  106 and 1.669  106 respectively. Moreover, the viscosities
of refined AGF at low concentrations (below 4.5 wt%) were lower
than that of refined ARF, while at concentration of 4.5 wt% (or
above), viscosities of both species were equivalent. These had
important implications for guiding the practical application of
refined KF. Furthermore, rheological properties of refined AGF were
susceptible to pH, especially at pH 2, giving rise to the potential
value as pH-sensitive material. Nevertheless, Properties of refined
ARF were nearly not influenced by the external environment,
which contributed to a wide range of application in various fields.
Furthermore, the thermo-gelling temperature of refined AGF was
lower than that of refined ARF, which would be a pronounced
advantage for A. guripingensis to be widely used in food industry.
Acknowledgments
This work was financially supported by Key Project of Chinese
Ministry of Education (Grant No. 113047A) and the National Natural
Science Foundation of China (Grant No. 31371841). The authors
expressed greatly thank to colleagues of Key Laboratory of
215
Environment Correlative Dietology of Huazhong Agricultural Uni-
versity for offering many conveniences.
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