LWT - Food Science and Technology 77 (2017) 92e99

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LWT - Food Science and Technology

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

Ultrasound assisted extraction of polysaccharides from mushroom

by-products

-Aguayo a, *, Jennie Walton b, Inmaculada Vin
Ingrid Aguilo ~ as c, Brijesh K. Tiwari d, **
a gic Agroalimentari de Lleida, Lleida 25003, Catalonia, Spain
IRTA, XaRTA-Postharvest, Edifici Fruitcentre, Parc Científic i Tecnolo
b
Manchester Food Research Centre, Manchester Metropolitan University, UK
c
Food Technology Department, University of Lleida, XaRTA-Postharvest, Agrotecnio Center, Rovira Roure 191, 25198 Lleida, Catalonia, Spain
d
Teagasc Food Research Centre, Dublin, Ireland

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

Article history: The effect of ultrasound technology to extract the water soluble polysaccharides from dried and milled
Received 22 July 2016 by-products generated from Agaricus Bisporus production was studied. Amounts of b-glucan 1.01 and
Received in revised form 0.98 g/100 g dry mass were obtained in particle sizes of 355e250 mm and 150-125 mm from the
20 October 2016
mushroom by-products. Three parameters of extraction were studied; extraction time (0e15 min), ul-
Accepted 14 November 2016
Available online 15 November 2016
trasonic amplitude (20e100 mm) and precipitation time (1 or 18 h). The application of ultrasounds
enhanced the extraction polysaccharide yields compared to the untreated samples. The highest extrac-
tion yield of 4.7% was achieved with an extraction time of 15 min, maximum amplitude of 100 mm with
Keywords:
Ultrasound
1 h of precipitation in 80% ethanol. The coefficient of determinations for predicted water soluble poly-
Water soluble polysaccharides saccharides extraction yields showed good correlation with the experimental data at the 95% confidence
Mushrooms level and indicated that the non-exponential Peleg's model could be employed to predict the extraction
Peleg's model polysaccharide yields after ultrasound treatment.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction Naczk, 2013). However, these methods require long extraction

High quantities of by-products are generated during white
mushroom (Agaricus Bisporus) cultivation. This material is made up
mainly of stalks and cups that are misshaped or do not meet the
specifications set by retailers, representing between 5 and 20% of
fruiting bodies discarded (Wu & Sun, 2010). They offer significant
economic potential as fungal cell walls containing chitin, other
hemicelluloses, mannans and, among the most interesting func-
tional components, b-glucans (Wong & Cheung, 2009). b-glucan is
one of the soluble dietary fibres that has been shown to enhance
immune function, lower blood cholesterol and potentially atten-
uate the glycaemic load of foods (Charlton et al., 2012; Shimizu
et al., 2008; Wolever et al., 2011).
Hot water and alkali extraction followed by precipitation with
alcohol are the most common techniques used for polysaccharides
extraction, including b-glucans (Lewis, 1984; Zhong, Shahidi, &
* Corresponding author.
** Corresponding author.

-Aguayo), Brijesh.Tiwari@
E-mail addresses: Ingrid.Aguilo@irta.cat (I. Aguilo
teagasc.ie (B.K. Tiwari).
times and high temperatures giving low extraction yields (Huang &
Ning, 2010). This has led to use novel extraction techniques
including ultrasound-assisted extraction (UAE), microwave-
assisted extraction and supercritical fluid extraction. Among
these, UAE offers an inexpensive, environmentally friendly, less
time consuming and efficient alternative to conventional extraction
techniques (Chavan & Singhal, 2013; Hossain et al., 2012). The
enhancement in extraction caused by ultrasound is mainly attrib-
uted to the effect of acoustic cavitations produced in the solvent by
the passage of an ultrasound wave (Ma et al., 2008; Veli ckovi
c,
Milenovi
c, Risti

c, & Veljkovi

c, 2008). Therefore, cell wall structure
is disrupted and diffusion through membranes is accelerated
(Benito-Roma
n, Alonso, & Cocero, 2013).
The application of ultrasound as a technique for assisting
extraction of bioactive plant origin metabolites (Knorr, 2003;
Vilkhu, Mawson, Simons, & Bates, 2008), flavonoids from foods
using a range of solvents (Zhang, Xu, & Shi, 2003) and bioactives
from herbs (Vinatoru, 2001) has been widely reported. Previous
studies have reported the feasibility of UAE to extract poly-
saccharides from A. Bisporus by optimizing parameter conditions by
applying orthogonal or central composite design experiments

http://dx.doi.org/10.1016/j.lwt.2016.11.043
0023-6438/© 2016 Elsevier Ltd. All rights reserved.

-Aguayo et al. / LWT - Food Science and Technology 77 (2017) 92e99
I. Aguilo
resulting in extracting polyssacharides yields of around 5e6 g/
100 g (Qiao, Zhao, Huang, Fan, & Han, 2012; Tian et al., 2012; Chen,
Zhou, Li, Bi, & Yang, 2014). However, other paramaters to optimize
the extraction polyssacharide yields could be taken into account
such as ultrasonic amplitude or precipitation time. On the other
hand, quadratic regression models have been used to describe UAE
of polysaccharides from fungal material (Tian et al., 2012; Yan et al.,
2016). The non-exponential Peleg's model has been successfully
used to describe extraction kinetics of various bioactives from
range of matrices such as UAE of anthocyanins from black choke-
berry (Galva
n, Dimitrov, Vauchel, & Nikov, 2014), solid-liquid
extraction kinetics of polyphenols from grape seeds (Buci
c-Koji
c,
Planini

c, Tomas, Bili

, 2007), ultrasound assisted rehydra-
c, & Velie
tion and dehydration processes (Clemente, Sanjuan, Carcel, &
Mulet, 2014; Ghafoor, Misra, Mahadevan, & Tiwari, 2014; Hui-
Xiao, Xi-Hua, Ming-Hui, Dong-Yang, & Yu-Jie, 2012) and for the
intensification of ursolic acid ultrasound extraction from Ocimum
sanctum (Vetal, Lade, & Rathod, 2013).
Therefore, the aim of this study was to study the application of
ultrasound technology to extract water soluble polysaccharides
(WSP) from dried by-products of A. Bisporus production. Optimi-
zation of extraction times and ultrasound amplitude on the
extraction yield of WSP after precipitation was also determined.
Moreover, the flexibility of Peleg model to provide key information
about extraction kinetics will be evaluated to predict different
extraction kinetic parameters. On the other hand, the impact of the
particle size of the dried material on the b-glucan levels, as pre-
dominant soluble dietary fibre, was evaluated.
2. Materials and methods
2.1. Sample preparation
Mushroom by-product materials were kindly provided by G's
Fresh at Chelbury mushroom farm (Cheltenham, UK). Around 15 kg
of mushroom material was donated, including mushroom cups and
stalks. Fresh mushrooms were also obtained from Sainsbury's Ltd
(UK). As the mushroom by-products contained a large amount of
soil residue, the material was washed in lukewarm water to remove
the soil. They were then dried using a hot air method at 60
C until
they reached a constant weight. Dried mushroom material were
then milled in ZM100 Mill (Retch, Dusselfdorf, Germany) in to a
powder with a particle size of less than 0.25 mm, vacuum packed
and stored at 4
C.
2.1.1. Sample preparation for the study on b-glucan content
In order to determine the effect of particle size on the b-glucan
content of the mushroom by-product powder, 100 g of sample was
separated according to their particle size using a sieve shaker
(Retch, Dusseldorf, Germany). The mesh size of the sieves were
>425, 355, 250, 150, 125 and 90 mm. The samples were placed in the
top sieve and shaken for 10 min at a setting of 60 rpm. The material
was collected together and put through the sieve shaker an addi-
tional 2 times. The remaining residue was weighed and expressed
as a percentage of the original sample weight. Each sample prep-
aration was done in triplicate.
2.2. Nutritional evaluation
2.2.1. General composition analysis
The general composition of the mushroom by-product powder
was determined using standard methods. Analysis of total moisture
content as well as soluble, insoluble and total fibre content was
done by following the Association of Official Analytical Chemists
(AOAC) methods (AOAC, 1995). Ash contents were determined by
93
burning in a muffle furnace at 550
C for 3 h according to the AACC
Approved Methods (AACC, 2000). Total fat content was determined
using the Caviezel method (Pendl, Bauer, Caviezel, & Schulthess,
1998). Protein contents were determined using a Kjeldahl diges-
tion system (KI 26, Gerhardt, Ko€nigswinter, Germany) based on the
AOAC methods. As mushrooms contain non-protein nitrogen
compounds, the conversion factor used to estimate protein content
was 4.7 (Mattila, Salo-Vaananen, Konko, Aro, & Jalava, 2002). Di-
etary fibre (total, insoluble, and soluble fraction was analysed by
the enzymatic-gravimetric AOAC method (Prosky, Asp, Furda, De
Vries, & Schweizer, 1988). Total carbohydrate was calculated as
follows (Eq. (1)):
Total carbohydrate ðg=100 g DMÞ
¼ 100  protein content ðg=100 g DMÞ
 lipid content ðg=100 g DMÞ  ash ðg=100 g DMÞ (1)
where DM was dry mass.
2.2.2. b-Glucan determinations
The content of b-glucan was determined enzymatically in the
different mushroom by-product powder fractions according to
their particle size (from >425 mm, 355-250 mm, 150-125 mm to
<90 mm) using the commercial Yeast Beta Glucan Assay Kit (Meg-
azyme International, Bray, Ireland). Total glucan was determined
first by solubilising the a-glucan and b-glucan linkages in hydro-
chloric acid, then potassium hydroxide (KOH) and then filtered.
Hydrolisation to D-glucose was then completed by the addition of a
mixture of exo-1,3-b-glucanase and b-glucosidase. Free Deglucose
was then measured spectrophotometrically at 510 nm a-Glucan
was then determined by solubilisation with KOH then hydrolysis to
D-glucose with amyloglucosidase plus invertase. The free D-glucose
was then measured spectrophotometrically at 510 nm. The b-
glucan content was calculated by inputting the data into Mega-Calc
™ excel sheet provided by Megazyme (Megaenzyme International
Ltd., Bray, Ireland).
Each analysis was performed in triplicate.
2.3. Ultrasound-assisted extraction of water soluble
polysaccharides
Ultrasound-assisted extraction (UAE) of WSP was carried out
following the method proposed by Chen et al. (2012), with some
modifications. An amount of 3 g of dried milled mushrooms were
weighed in to centrifuge tubes and 30 mL of distilled water was
added (1:10 w/v). Sonication was conducted by means of the
UP400S Ultrasonic Processor (maximum nominal power of 400 W,
24 kHz; Hielscher, Teltow, Germany) at differing ultrasound
amplitude levels (20, 60 and 100 mm amplitude) for varying periods
of time (0, 3, 5, 10 and 15 min). After sonication, the extract was
centrifuged at 12 000 g for 20 min at room temperature and the
residue and supernatant separated. The supernatant for each
sample was mixed with 80% ethanol (1:2 v/v) at room temperature.
This treatment was carried out in two conditions, one set of sam-
ples were left for 1 h and the other set were left for 18 h at 4
C.
Separation of the crude WSP precipitate was done by centrifugation
at 12 000 g for 20 min. The WSP was dried at 40
C and then
weighed. Yield was calculated using the following calculation (Eq.
(2)):
mt
Yðg=100gÞ ¼  100 (2)
mi
where mt was the crude extract and mi was the mass of the
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I. Aguilo

Table 1 experimental values. RMSE was estimated as following (Eq. (4)):

Nutritional composition of mushroom by-product powder.
" #1=2
Nutrients (g/100 g DMa) 1 XN
RMSE ¼ ðexperimental  calculatedÞ2 (4)
Ash 10.12 ± 1.04 N i¼1
Moisture 7.26 ± 0.05
Protein 26.16 ± 0.49
Crude lipids 3.39 ± 0.39
Carbohydrates 53.07 ± 0.68
Insoluble fibre 35.81 ± 4.97
3. Results and discussion
soluble fibre 1.42 ± 0.59
a-Glucan 0.12 ± 0.02
b-Glucan 0.98 ± 0.02 Nutritional composition of mushroom by-product powder was
Total Glucan 1.09 ± 0.02 determined and results were expressed on dry mass (DM) (Table 1).
Values are mean(n ¼ 3) ± Standard Deviation. The level of protein in waste mushroom was 26.16 g/100 g DM,
a
DM is dry mass basis. which was in agreement with the findings reported in literature in
edible mushrooms ranging between 23.9 and 34.8 g/100 g DM
(Chang & Miles, 2004; Cheung, 2008). The moisture content (7.26 g/
mushroom powder. 100 g DM) was lower than values of moisture content reported in
fresh edible mushrooms (between 60 and 90 g/100 g) (Hung & Nhi,
2.4. Kinetic of ultrasound-assisted extraction 2012). This variability could be dependent on the mushroom spe-
cies and other parameters such as environmental temperature,
A non-exponential empirical equation (Eq. (3)) was used to relative humidity during growth and relative of metabolic water
model the extraction yield of WSP from mushroom by-product. that may be produced or used during storage (Zaki, El-Kattan,
Peleg (1988) proposed the same kinetic model equation (Eq (3)) Hussein, & Khaled, 1993). Lipid contents of mushroom by-
for sorption curves. products were found to be 3.39 g/100 g of DM. Lipid content
could vary from 1.3 g/100 ge8.0 g/100 g on DM basis depending on
t the nutritive of substrate used to grow mushrooms, genus of spe-
Ct ¼ C0 þ (3)
K1 þ K2 t cies, environment conditions (Crisan & Sands, 1968; Hung & Nhi,
2012; Zaki et al., 1993). The ash content (10.12 g/100 g DM)
where Ct was the yield of WSP at time t (g/100 g DM), C0 was the observed in these mushroom by-products revealed that these
initial yield of WSP (g/100 g DM), t was the extraction time (min), waste could be a good source of minerals including magnesium,
K1 was Peleg's rate constant (min 100 g/g DM) and K2 related to calcium, sodium, selenium, potassium, copper, iron and zinc (Chang
maximum (or minimum) attainable moisture content (100 g/g). & Miles, 2004). On a dried mass basis, the mushroom by-product
powder contained an average of 35% insoluble fibre and 1.42 g/
2.5. Statistical analysis 100 g DM soluble fibre (Table 1). Literature on the dietary fibre (DF)
content of A. Bisporus is limited, however the literature that has
Minitab version 15 (State College, Palo Alto, USA) was used for been found suggests that on average A. Bisporus contains 1.98 g/
data analysing. Data was analysed by analysis of variance (ANOVA) 100 g edible mass total dietary fibre (Manzi, Aguzzi, & Pizzoferrato,
and means separation was achieved using Tukey multiple range 2001) and 8e10 g/100 on DM basis (Chang & Miles, 2004; Cheung,
test with the significance level of 0.05. The root mean square error 2008). Total glucan content was 1.09 g/100 g DM, where 0.98 and
(RMSE) was used to determine the quality of the fit of Peleg's model 0.12 g/100 g corresponded to b and a-glucan amounts, respec-
by comparing the predicted extraction yield of WSP to the tively. Hammond (1979) found that after 4 d of storage, the chitin

40
By-product Quantity (g/100 g DM)

35

30

25

20

15

10

0
>425 425-355 355-250 250-150 150-125 125-90 <90
Particle size ( m)

Fig. 1. Particle size analysis of mushroom by-product powder.


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I. Aguilo 95

1.04

1.02

1.00

β-Glucan content (g/100g DM)

0.98

0.96

0.94

0.92

0.90

0.88
> 450 355-250 150-125 <90
Parcle size ( m)
Fig. 2. b-Glucan content (g/100 g DM) of various fractions of mushroom by-product powder.

(insoluble cell wall component classed as DF) content of A. Bisporus
increased by over 50%. This effect may lead to varying results as the
mushrooms used in literature were assumed to be fresh whereas
the present study used waste mushrooms that were kept for over
4 d.
Total carbohydrates including polysaccharides such as glucans,
mono- and disaccharides, sugar alcohols, glycogen and chitin were
of 53.07 g/100 g DM, which was found to be in the same range than
those values reported on other edible mushroom species (Hung &
Nhi, 2012; Zaki et al., 1993).
mushroom by-product powder at precipitation times of 1 h and
18 h is shown in Fig. 2a and b, respectively. In general, the appli-
cation of ultrasounds enhanced the extraction polysaccharide
yields compared to the untreated samples. Maximum WSP
extraction yields of 4.70 and 4.35 g/100 g DM were obtained at
amplitudes of 100 mm and treatment times for 15 min at precipi-
tation times of 1 h (Figs. 2a) and 18 h (Fig. 2b), respectively. The
application of ultrasound technology was previously used in the

3.1. Effect of particle size separation on b-glucan

The analysis of the particle size of the mushroom by-product

powder indicated that the by-product quantities of 35.26 and
21.47 g/100 g DM corresponded to particle sizes of <90 mm and
between 250 and 150 mm, respectively (Fig. 1). Only 2.91 g/100 g of
DM waste quantity had a particle size higher than 450 mm. Prior to
particle size separation, b-glucan content was characterized in
waste mushroom powder indicating an amount of 0.997 g/100 g on
DM basis (Table 1). Manzi et al. (2001) reported that content of
A. Bisporus was 0.14 g/100 g based on fresh mass. b-glucan per-
centages were determined in the fractions of >450 mm, 355-
250 mm, 150-125 mm and <90 mm in order to observe the influence
of particle size on the b-glucan content (Fig. 2). The highest b-
glucan amounts of 1.01 and 0.98 g/100 g DM were observed in
particle sizes of 355e250 mm and 150-125 mm, respectively (1.01 g/
100 g DM). Although smaller particles may contain higher contents
of b-glucan, during extraction it could not be lead to larger amounts
of extracted polysaccharide. A study conducted on the extraction of
polysaccharides from BaChu mushroom by XuJie and Wei (2008)
indicated that smaller particle sizes resulted in lower extraction
yields than higher particle sizes. XuJie and Wei (2008) suggested
that this was due to a decrease resistance and a shorter path for
diffusion leading to a smaller amount of polysaccharide moving
from the interior of the particle in to the solvent.

3.2. Ultrasound-assisted ectraction of water soluble

polysaccharides

The effect of ultrasound power levels and treatment times on Fig. 3. WSP extraction yield (g/100 g DM) after a) 1 h and b) 18 h precipitation time.
the extraction yield of water soluble polysaccharides (WSP) from Ultrasound power levels at ( ) 20, (,) 60 and at (D) 100 mm amplitude.
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I. Aguilo
extraction of polysaccharides from fungal material (Chen et al.,
2012; Tian et al., 2012; Yan et al., 2011) enhancing extractability
by using lower extraction times than conventional methods (Guo,
Zou, & Sun, 2010; XuJie & Chen, 2008). Structural changes caused
by ultrasonic cavitation enhanced the extraction of target com-
pounds from the material to the surroundings by increasing
microfractures on the surface of samples (Li, Pordesimo, & Wiess,
2004; Suslick & Price, 1999).
In order to better visualise the relationship between the ultra-
sonic treatment time and amplitude on the WSP extraction yields,
3-D response surface graphs were also plotted as function of pre-
cipitation time (Fig. 3). No significant difference between extraction
yields (P>0.05) were observed as function of 1 or 18 h of precipi-
tation time. Extraction yield rose when only increasing ultrasound
treatment time from 0 to 20 min, irrespective of precipitation time.
Increasing the extraction time allows the solvent to be in contact
with the material for a longer period of time, allowing higher
diffusion rates. However, if the extraction time is extended too
much the yield can begin to slowly decrease due to insoluble ma-
terial also being suspended in the solvent after rupturing of the
Fig. 4. Response surface plot for WSP extraction yield (g/100 g) after a) 1 h and b) 18 h holding time.
cells, which will lower the permeability of the solvent (Maran,
Manikandan, Nivetha, & Dinesh, 2013). On the other hand, an in-
crease in ultrasound amplitudes also led to higher polysaccharide
extraction yields, irrespective of the precipitation time (Fig. 3). An
increment in the applied ultrasound power could lead to an in-
crease in polysaccharide extraction due to a more aggressive
collapse of cavitational bubbles caused by the increment of the
resonant bubble size proportional to the amplitude of the ultra-
sound wave (Li, Pordesimo and Wiess, (2004); Maran et al.,2013)
(see Fig. 4).
The combined effect of both ultrasound treatment time and
amplitude showed a positive influence on the WSP extraction from
mushroom by-product powders, meaning that longer and intense
ultrasound treatments could lead to higher extraction yields
(Fig. 3). These results agreed with those reported by Tian et al.
(2012) on the extraction of WSP from A. Bisporus, with an in-
crease in the extraction yields when rising ultrasound powers from
200 W to 230 W. However, Chen et al. (2012) reported that
decreasing extraction times from 10 to 8 min led to increasing
yields in the extraction of WSP from the mushroom species Boletus

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I. Aguilo 97

Edulis mycelia. Differences in mushroom varieties may explain the precipitation was carried out by Kozarski et al. (2011). They could
differences in the optimal ultrasound extraction conditions. How- extract a total polysaccharide content of 74.4 g/100 g dry mass
ever, these studies also included other variables such as the tem- exclusively extracted from fresh mushrooms fruit boodies cap.
perature and W/M. Tian et al. (2012) reported maximum extraction Higher solvent to solid ratio could induce a concentration dif-
yields of 6.02 g/100 g (with protein removed) at 230 W, extraction ference between the exterior solvent and the interior cells of the
times of 62 min, temperatures of 70
C and a W/M ratio of 30 mL/g. solid, creating favourable conditions for mass transfer. In the same
Similar extraction yields of 5.6 g/100 g from A. bisporus were ach- way, higher temperatures could increase mass transfer by acceler-
ieved by Qiao et al. (2012) by applying extracting temperatures of ating molecular movement, leading to an increase in the solubility
65
C, extraction time of 40 min, W/M of 30 mL/g with an ultrasonic of material (Maran et al.,2013). In the current study, room tem-
power of 170 W. In contrast, Chen et al. (2012) observed maximum peratures and constant ratios of 1:10 w/v have been kept constant
extraction yields of 15.48 g/100 g from the same matrix when in the experimental design. Although higher temperatures could
applying temperatures of 56
C, 1:55 of ration of dried mycelia to increase extraction yields, the purity of the extract might be
water for 8.4 min. Yan et al. (2011) reported the highest poly- affected. Li et al., 2004 obtained polysaccharide yields of 20.2 g/
ssacharides yield of 8.26 g/100 g from Tremella mesenterica at 100 g from Zizyphus jujuba cv. jinsixiaozao when applying temper-
extraction time of 30 min at 50
C and W/M of 34 mL/g. There are atures of 45e53
C, sonic powders of 31.7 W, and treatment times
several reports describing the determination of polysaccharides of 20 min and water/solid ration of 20:1. However, the chemical
from A. bisporus by other extraction techniques. Yin, You, and Zhou analysis of crude polysaccharides revealed the polysaccharides
(2015) extracted polyssacharides from A. bisporus by complex extracted were most contaminated by co-extracted proteins.
enzyme-assisted extraction with yields of 6.87 g/100 g. Xue and
Farid (2015) achieved polysaccharide extraction values of 0.79 g/ 3.3. Ultrasound-assisted extraction of water soluble
100 g mushroom by applying pulsed electric fields at intensities of polysaccharides
38.4 kV/cm for 272 ms and a treatment temperature of 85
C.
Parniakov, Lebovka, Heche, and Vorobiev (2014) compared the ef- WSP ultrasound extraction curves (Fig. 3) showed similarity
ficiency of extraction and stability of extracts for different methods with course of sorption processes, which could be well described by
of extraction including pressure extraction, pressure extraction the non-exponential Peleg's model (1988). The experimental WSP
assisted by pulsed electric field (PE þ PEF), hot water extraction, extraction yields, calculated parameters of Peleg's model (constants
ethanol extraction and supplementary ethanol extraction from K1 and K2), regression coefficient (R2) and RMSE are shown in
cakes of slices. They observed that PE þ PEF allowed production of Table 2. The model fitted the experimental data as demonstrated
mushroom extracts with the highest polysaccharides content the high significance of the model (P < 0.01) and high R2 values
ranging 3 g/100 g dry matter. Partially purified polysaccharides between 0.627 and 0.995 which implied good concordance be-
from A. bisporus by hot water extraction followed by ethanol tween experimental and calculated data. The RMSE had low values

Table 2
Values of WSP extraction yield, Peleg's constants (K1 and K2) for ultrasound extraction with regression coefficient (R2) and the root mean square error (RMSE) at different
extraction time and amplitude.

Extraction time (h) Amplitude (mm) WSP extraction yield (g/100 g DW) K1 (min$g/100 g) K2 (100 g/g) R2 RMSE

1 20 2.23 ± 0.36 0.73 ± 0.03 1.04 ± 1.83 0.995 0.090

60 2.76 ± 0.03 0.90 ± 0.21 1.60 ± 1.04 0.700 0.417
100 3.10 ± 0.21 0.78 ± 0.04 1.09 ± 1.76 0.627 0.162
18 20 2.78 ± 0.14 1.49 ± 0.25 1.57 ± 1.59 0.809 0.299
60 2.57 ± 0.01 0.83 ± 0.25 2.72 ± 1.77 0.961 0.256
100 2.800.13 0.39 ± 0.06 2.90 ± 1.30 0.875 0.370

a) b)
5.0
5.0
Predicted WSP extracƟon yield

Predicted WSP extracƟon yield

4.5
4.5
4.0
4.0
3.5

3.0 3.5

2.5 3.0

2.0 2.5
1.5
2.0
2.0 2.5 3.0 3.5 4.0 4.5 2.5 3.0 3.5 4.0 4.5 5.0

A c tual WSP e xtr ac tion yie ld A c tual W SP e xtr ac tion yie ld

Fig. 5. Predicted and experimental values for Peleg model fitted to WSP extraction yield after one hour of extraction (a); after 18 h of extraction (b). — 95% PI, —95% CI.
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I. Aguilo
for extraction times of 1 h and ultrasound amplitudes of 20 mm. The
extraction rate constant (K1) showed a tendency to decrease with
the increase of ultrasound amplitudes when using extraction times
of 18 h. To determine the accuracy of the predicted model, a set of
data from additional test runs were obtained. Predicted values of
WSP extraction yields obtained using Peleg's equations were in
good agreement with the experimental values (Fig. 5). The exper-
imental and predicted values closely correlated, as demonstrated
by R2 values of 0.93 and 0.87 for 1 and 18 h of precipitation,
respectively. The predicted values were within the 95% confidence
interval, as illustrated in Fig. 5. Therefore, the predictive perfor-
mance of the established model may be considered acceptable
under the experimental conditions employed.
4. Conclusions
The b-glucan content of the by-products of A. Bisporus produc-
tion was studied. The highest b-glucan amounts of 1.01 and 0.98 g/
100 g DM were observed in particle sizes of 355e250 mm and 150-
125 mm, respectively. Thus, indicating that particle size had no
significant effect on the b-glucan content.
The effect of ultrasound technology on the extractability of
water soluble polysaccharides from by-products of A. Bisporus
production was studied. The results indicated that increasing
extraction time and ultrasonic amplitude led to greater extraction
yields. Maximum extraction yields of 4.70 and 4.35% were obtained
after applying ultrasonic amplitudes of 100 mm and treatment
times for 15 min at precipitation times of 1 and 18 h, respectively.
Results showed that the efficiency of the extraction treatment
could be improved by reducing the precipitation time from 18 h to
1 h as no significant effect on the extraction yield was observed.
Moreover, the finding suggests that Peleg's model could be
employed to describe the WSP extraction yield during ultrasound
treatment.
Acknowledgements

-Aguayo thanks to the Ministry of Economy and
Dr. Aguilo
Competitiveness from the Government of Spain for the “Ayuda para

n postdoctoral” (FPDI-2013-15583).
contratos para la formacio
References
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