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Flaxseed Gum From Flaxseed Hulls
Flaxseed Gum From Flaxseed Hulls
Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd
University of Guelph, Department of Food Science, Guelph, ON, Canada N1G 2W1
Agriculture and Agri-Food Canada, Guelph Food Research Centre, 93 Stone Rd. W., Guelph, ON, Canada N1G 5C9
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 24 September 2011
Accepted 21 December 2011
Soluble dietary bre with low viscosity has the potential to deliver acceptable mouthfeel and texture
when included in the diet in a signicant amount to show health benets. Soluble axseed gum (SFG)
was reported to have low viscosity, thus has potential in applications as a bre fortier. In the present
work, SFG extracted from axseed hulls was fractionated into a neutral fraction gum (NFG) and an acidic
fraction gum (AFG) using ion exchange chromatography. The protein content in SFG (11.8%) and AFG
(8.1%) were completely removed by protease to obtain two more protein-free fractions, SFGnP and
AFGnP; NFG contained no protein. The uronic acid content in NFG (1.8%) was mostly eliminated, whereas
in AFC increased to 38.7%. NFG consisted of high molecular weight (MW) (1470 kDa) arabinoxylans and
exhibited pseudoplastic ow behaviour; whereas AFG was mainly composed of rhamnogalacturonans
with a higher MW fraction (1510 kDa) and a lower MW fraction (341 kDa) and showed Newtonian ow
behaviour. The ranking of intrinsic viscosities (mL g1) in decreasing order was: SFG (446.0) > NFG
(377.5) > AFG (332.5). AFG was expected to have higher chain exibility for its lower value of Huggins
constant (0.16) compared to that of NFG (0.54) and SFG (0.48). In comparison with SFGnP, AFGnP and
NFG, SFG and AFG showed better surface activities and emulsifying stabilities due to the presence of
protein in both fractions.
Crown Copyright 2012 Published by Elsevier Ltd. All rights reserved.
Keywords:
Flaxseed gum
Fractionation
Rheology
Intrinsic viscosity
Critical concentration
Emulsifying properties
1. Introduction
Flax (Linum usitatissiumum L.) was rst introduced to North
America as a crop for structural bres, but its value and importance
as an oil source quickly became apparent (Cunnane & Thompson,
1995). Canada is the leading country in producing and exporting
oil-type axseed. Western Canadian axseed is composed of 45%
oil and 23% protein (1990e2008 means) (www.grainscanada.gc.ca).
The renewed interest in axseed as a food source is due to its health
benets attributed to its components including lignans (secoisolariciresinol diglucoside (SDG) being the predominant form), alinolenic acid, and soluble axseed gum (SFG) (Hall Iii, Tulbek, & Xu,
2006). In comparison to locust bean gum, guar gum and xanthan
gum at a concentration of 0.3% (w/v), SFG exhibited much lower
viscosity over a range of shear rate from 10 to 1000 S1 (Mazza &
Biliaderis, 1989). Its low viscosity is favoured in dietary bre fortication in food without leading to an over-texturization, when
a signicant concentration of bre is required to show health
benets.
* Corresponding author. Tel.: 1 519 780 8028; fax: 1 519 829 2600.
E-mail addresses: kqian@uoguelph.ca (K.Y. Qian), cuis@agr.gc.ca (S.W. Cui),
Wuy@agr.gc.ca (Y. Wu), dgoff@uoguelph.ca (H.D. Goff).
0268-005X/$ e see front matter Crown Copyright 2012 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodhyd.2011.12.019
276
Fig. 1. Flow chart for extraction, fractionation and characterization of soluble axseed gum (SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction gum; IEC: ion
exchange chromatography; Buffer 1: 20 mM Tris/HCl, pH 8).
hrel h=hs
(1)
hsp hrel 1
(2)
(3)
(4)
277
the solution and solvent (MilliQ H2O); hrel and hsp are two dimensionless parameters of relative and specic viscosity; and hred and
hinh represent reduced viscosity and inherent viscosity, respectively. KH (generally positive) and KK (generally negative) are
Huggins and Kraemer constants in Huggins and Kraemer equations
(Eqs. (3) and (4)). Both equations are valid only for solution
viscosity up to twice of the solvent viscosity (i.e. hrel 2). Beyond
this region, with concentration increase polymerepolymer interactions will progressively become signicant, thus higher-order
terms (c2, c3, etc.) will no longer be negligible. A lower limit of
hrel 1.2 is also required to eliminate the increasing errors in hsp as
hsp approaches 0. Concentrations of each gum fraction adopted
were conned to this region (1.2 hrel 2). The value of intrinsic
viscosity ([h]) was reported as the mean of both intercepts in linear
functions (3) and (4) by plotting hsp/c and lnhrel/c against c and
extrapolating each linear trendline to zero concentration.
2.5. Functional properties
2.5.1. Surface activity
The surface tension of the airewater interface was measured by
the Du Nouy ring method using the Fisher Surface Tensiomat
(model 21, Fisher Scientic, Nepean, ON) (Izydorczyk, Biliaderis, &
Bushuk, 1991). SFG and its fractions were dissolved in 20 mL of
distilled water at various concentrations, heated at 80 C for 1 h
under gentle stirring and then left unperturbed for 2 h before
measurements. The surface tension (dyn cm1) of distilled water
and gum solutions were measured every 5 min for 6 times at 25 C.
2.5.2. Emulsifying properties
Emulsions were prepared by mixing canola oil (2% wt) with SFG
or its fraction dispersions (preheated at 80 C for 1 h, nal gum
concentrations were adjusted to 0.44% wt) using Polyton (Kinematica GmbH, Brinkmann homogenizer, Switzerland) at medium
speed (level 5) for 5 min, followed by homogenization (Nano DeBEE
Electric Bench-top Lab Homogenizer, BEE International, South
Easton, MA, USA) with 2 passes at 5000 psi.
The particle size distribution of the emulsions was measured
using integrated light scattering (Mastersizer X, Malvern Southborough, MA) (Khallou et al., 2008). The measurements were
performed within one day after emulsion preparation as phase
separation occurred in all the emulsions within 24 h. The emulsifying stability was determined by comparing the shifts of the oil
droplet size distributions.
3. Results and discussions
3.1. Extraction, fractionation and chemical composition
The composition and yield of axseed gum has been reported to
vary with extraction conditions, as well as culture environment and
genotype. Gum yield from axseed increased from 4 to 9.4 % with
extraction temperature increased from 25 to 80 C (Fedeniuk &
Biliaderis, 1994). Broad variations of 3.6e8.0% among 109
cultivars (80 C, 2 h) and 5.4e7.9% among 12 genotypes (85 C,
3 h) were also found by Cui, Kenaschuk, and Mazza (1996) and
Oomah et al. (1995), respectively. Soaking axseed with a water:
seed ratio of 13 at 85e95 C and pH from 6.5 to 7.0 for 3 h was found
to be the optimum gum extraction condition to achieve higher yield
and quality, using response surface methodology (Cui, Mazza,
Oomah, & Biliaderis, 1994). However, higher extraction temperature induced higher protein content and browning in colour
(Fedeniuk & Biliaderis, 1994). Considering the browning may affect
the properties of the polysaccharides, in the present work, the
extraction of the gum was conducted at room temperature.
278
The yield and chemical composition of SFG, NFG and AFG are
shown in Table 1. The yield of SFG is 9.7% of the hull mass. The
recovery of the chromatographic fractionation was approximately
50%; the loss of half of the SFG may partially be due to physical
blockage of large particles in the column and/or insufcient
unbinding process while washing AFG out. The uronic acid content
in AFG was about 20 fold of that in NFG, suggesting an efcient
separation of NFG from AFG. NFG was free of protein, while AFG
(8.1%) contained less protein than SFG (11.8%). After protein
removal from SFG and AFG, SFGnP and AFGnP were obtained with
their protein content decreased to an undetectable level.
Relative neutral monosaccharide composition of SFG and its
fractions is shown in Table 2. No signicant differences were found
between SFG and SFGnP, implying the removal of protein via
enzymatic hydrolysis from SFG fraction did not modify the neutral
sugar composition. However, the same enzymatic hydrolysis
procedure changed the three main components in AFG: small
portion of Rhamnose was possibly removed causing the relative
increase in the other two (Galactose and Fucose). These changes in
SFG and SFGnP were not signicant, since AFG occupied only w27%
of SFG. The neutral monosaccharide composition of SFG, NFG and
AFG was comparable with previous results (Cui, Mazza, & Biliaderis,
1994; Fedeniuk and Biliaderis, 1994). NFG was mainly composed of
xylose (68.2%) and arabinose (20.2%) therefore being identied as
arabinoxylans (Warrand et al., 2003). Its minor components
included galactose (7.9%) and glucose (3.7%), but no rhamnose or
fucose. In contrast, AFG mainly consisted of rhamnose (38.3%),
galactose (35.2%) and fucose (14.7%). With most galactose and all
fucose existing as terminals in side chains (Naran et al., 2008), AFG
was referred to as rhamonogalacturonans for its high content of
rhamnose and galacturonic acid (38.7%, Table 1.). The presence of
small amount of xylose (8.9%) and arabinose (2.9%) in AFG might be
due to the residual fraction of NFG (Cui, Mazza, & Biliaderis, 1994), or
might be derived from side chain components covalently linked to
its backbone (Naran et al., 2008).
Yield
Protein
Uronic acid
NFG
a
9.7 0.3
11.8 1.0
23.0 0.1
23.2 0.9
nd
1.8 1.0
AFG
b
SFG
Fucose
Rhamnose
Arabinose
Galactose
Glucose
Xylose
27.3 3.0
8.1 0.4
38.7 1.0
SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction gum; nd: not
determined; all data were calculated on a dry basis.
a
yield was based on axseed hull mass.
b
yield was based on SFG mass.
7.0
16.5
12.7
22.4
2.7
38.6
SFGnP
0.2c
0.6c
0.1b
1.0c
0.1b
1.2b
7.0
14.5
12.6
22.5
3.1
40.9
0.4c
0.3c
0.5b
0.7c
0.3ab
2.1b
NFG
AFG
AFGnP
nd
nd
20.2 0.1a
7.9 0.1d
3.7 0.4a
68.2 0.6a
14.7 0.3b
38.3 2.3a
2.9 0.4c
35.2 0.4b
nd
8.9 1.3c
16.5 0.1a
32.8 0.7b
3.0 0.1c
39.7 0.4a
nd
7.9 0.3c
SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction gum; data are
given as mean SD, n 3; data in the same row followed with the same
superscript letter are not signicantly different by Duncans multiple-range test; nd:
not determined.
h KM a
Table 1
Yield and chemical components of axseed gum and its fractions (%).
SFG
Table 2
Neutral monosaccharide in axseed gum and its fractions (%).
(5)
279
Fig. 2. Molecular weight (DRI) distributions of polysaccharides, column temperature 40 C (a); and their protein (UV) fractions, column temperature, 40 C (b) (SFG: soluble
axseed gum; NFG and AFG: neutral and acidic fraction gum; the unit of numbers in (a) is KDa).
h hN SI 0:5
(6)
y
S B$ hI0:1
(7)
Fig. 3. The steady shear ow curves (a) and dynamic rheological properties (b) of SFG, NFG and AFG solutions (SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction
gum).
280
Table 3
The intrinsic viscosity ([h]), Huggins constant (KH), critical concentrations (ccr) and
coil overlap parameter values ((c[h])cr) at critical concentrations of SFG, NFG and
AFG.
[h]
KH
ccr
(c[h])cr
(mL g1)
/
(g L1)
/
SFG
NFG
AFG
446.0 3.4
0.48
6.61
2.95
377.5 5.7
0.54
6.68
2.52
332.5 6.8
0.16
6.43
2.14
SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction gum.
hsp k$c$hn
(8)
values of (c[h])cr fell within a range from 2.14 to 2.95, not far away
from the values reported between 2.5 and 4 (Morris et al., 1981).
The values of n1 were conned to 1.08e1.21 (Fig. 4) and were
comparable with the value (1.1e1.4) reported (Doyle et al., 2009;
Morris et al., 1981; Wang & Cui, 2005). The n2 for SFG (3.28) and
NFG (3.51) were close to the typical value (w3.3) for various
disordered polysaccharides (Morris et al., 1981), though higher
value (n2 w5) was also found in the chain of several galactomannans (Doyle et al., 2009) due to the hyperentanglement
of unsubstituted mannan regions. Compared with SFG and AFG,
a much lower value of n2 (2.17) detected for AFG may probably also
be due to its relatively higher exibility and thus be less likely to
form an entanglement network, just as implied from its lower
value of KH;AFG (0.16).
Fig. 4. The intrinsic viscosity (a) and the dependence of viscosity on concentration (bed) of SFG, NFG and AFG (SFG: soluble axseed gum; NFG and AFG: neutral and acidic fraction
gum; viscosity was measured in 0.1 M NaCl (aed). Reduced viscosity (hred hsp/c) and inherent viscosity (hrel (lnhrel)/c) versus concentration are plotted with empty and solid
symbols (a), respectively; the common intercept gives [h], and the positive and negative slopes are KH[h]2 and KK[h]2, where KH and KK are Huggins and kraemer constant,
respectively (a). n1 and n2 represent the slope of each region; the gum concentration for the circles within the squares in gure (bed) equals to 5 g L1).
S u rfa c e te n s io n
(d y n e s /c m )
75
D4; 3
X
i1
70
NFG
65
AFGnP
60
SFGnP
55
AFG
SFG
50
0
0.1
0.2
0.3
0.4
0.5
ni d4i =
281
X
i1
ni d3i
fi di
(9)
i1
Fig. 6. The shifts of D[4,3] and particle size distribution proles of emulsions stabilized by SFG and its fractions within 24 h after emulsication with 2% wt of canola oil (SFG and
SFGnP: soluble axseed gum with and without protein; NFG: neutral fraction gum; AFG and AFGnP: acidic fraction gum with and without protein).
282
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