Food Chemistry 114 (2009) 623628

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Characteristics of axseed hull oil q

B. Dave Oomah a,*, Laurie Sitter b
a

National Bioproducts and Bioprocesses Program, Pacic Agri-Food Research Centre, Agriculture and Agri-Food Canada, 4200 Highway 97,
P.O. Box 5000, Summerland, BC, Canada V0H 1Z0
Universit de Bretagne Occidentale, Institut Universitaire Professionnalis, 29000 Quimper, France

a r t i c l e

i n f o

Article history:
Received 17 July 2008
Received in revised form 22 August 2008
Accepted 30 September 2008

Keywords:
Hull oil
Lipid fractions
DSC
Antioxidant activity
Solvent extraction
Supercritical CO2
SDG
Flaxseed

a b s t r a c t
Oils from two commercial axseed hulls extracted by six procedures were evaluated for physicochemical
characteristics. Oil yield ranged from 9% to 28% depending on solvent and extraction. Lipid fractionation
of crude axseed hull oil yielded 92.5% neutral lipids, 3.1% phospholipids, 2.4% acidic lipids and 2.1% free
fatty acids. Flaxseed hull oil exhibited three thermal transitions between 35 and 13 C with solvent
dependent polymorphism. Thermal oxidation by differential scanning calorimetry (DSC) revealed three
step oxidation of axseed hull oil with mean onset and oxidation temperatures at 121 and 150
253 C, respectively depending on the extraction procedure. Flaxseed hull oil exhibited two-fold difference (0.61.2 lm Trolox equivalent/g) in antioxidant activity measured by a photochemiluminescence
(PCL) assay. Supercritical CO2 extracted the most oil with the highest antioxidant capacity of all evaluated
procedures resulting in a defatted axseed hull containing the highest (53 mg/g) secoisolariciresinol
diglucoside (SDG) level.
2008 B. D. Oomah and L. Sitter of the department of Agriculture and Food, Government of Canada.
Published by Elsevier Ltd. All rights reserved.

1. Introduction
Recent studies have highlighted the numerous health benets
of axseed oil for the cardiovascular and skeletal systems (Griel
et al., 2007) and in inammatory conditions such as rheumatoid
arthritis, psoriasis, and ulcerative colitis (Mantzioris, James, Gibson, & Cleland, 1994). It can lower blood pressure particularly in
middle age dislipidaemic men (Paschos, Magkos, Panagiotakos,
Volteas, & Zampelas, 2007), and provides signicant improvement
in attention decit hyperactivity disorder in children (Stark, Crawford, & Reifen, 2008 and references therein). Flaxseed oil consumption has signicant effect on slowing bleeding time thereby
reducing the risk of myocardial infarction in type 2 diabetic Canadian male patients (Barre, Griscti, Mizier-Barre, & Hafez, 2005). In a
randomised controlled study, consumption of axseed oil signicantly increased the n3 (a-linolenic acid, eicosapentaenoic acid
and docosapentaenoic acid) fatty acid content in red blood cells
and in all tissues except brain (Barcel-Coblijn, 2007). Although
the demand for axseed oil as the consumers preferred source of
a-linolenic acid is being increasingly recognised by industry, regulators and consumers, due to heightened prole of omega-3 health
benets, a good reserve of this oil from axseed hulls remains untapped and underdeveloped.
q

Pacic Agri-Food Research Centre.

* Corresponding author. Tel.: +250 494 6399; fax: +250 494 0755.
E-mail address: oomahd@agr.gc.ca (B.D. Oomah).

Flaxseed hulls, a low-valued co-product obtained from ax

processing represents a potential source of value-added healthy
products. The hull, including the seed coat and endosperm, constitutes 36% of the total weight of hand-dissected axseed or 22% of
the seed when obtained mechanically (Oomah & Mazza, 1998).
Flaxseed hull is difcult to digest and therefore hinders access to
the lipids (Barcel-Coblijn, 2007). Oil content of axseed hulls varies from 26% to 30% depending on processing conditions (Oomah,
2003) representing approximately 18% of the total seed oil. This
oil from hulls obtained by dry abrasive dehulling contained significantly higher levels of palmitic acid and lowest level of stearic and
oleic acids compared to those from the whole seed (Oomah &
Mazza, 1997). Information currently available on characteristics
of axseed hull oil is decient and full investigations on oils from
commercially available axseed hull are long overdue. This
investigation describes the characteristics of oil obtained by
various solvents from axseed hulls to increase its value and contribution to the development of new omega-3 products for the
functional foods and nutraceutical applications.

2. Materials and methods

Flaxseed hulls were obtained from commercial sources
(Natunola Omega-3 ax hull from Natunola Health, Nepean, ON
and Pizzeys FortiGradTM from Pizzeys Milling, Angusville, MB).
Hulls from both sources were ground in a coffee grinder prior to

0308-8146/$ - see front matter 2008 B. D. Oomah and L. Sitter of the department of Agriculture and Food, Government of Canada. Published by Elsevier Ltd. All rights
reserved.
doi:10.1016/j.foodchem.2008.09.096

624

B.D. Oomah, L. Sitter / Food Chemistry 114 (2009) 623628

oil extraction by six different procedures. Oil from all hull samples
was extracted using hexane (50 g sample in 500 ml hexane) or
other solvents, as described by Oomah, Mazza, and Przybylski
(1996), purged with nitrogen and stored at 20 C until analysis.
Petroleum ether extracted oil was obtained using the Goldsh fat
extraction apparatus (Labconco, USA) for 6 h according to AOAC
International method (2000). Acetone oil was obtained by extracting hulls (50 g sample in 500 ml 50% aqueous acetone) for 15 h at
ambient temperature and the solvent removed by vacuum ltration. The residue was re-extracted with 100% acetone for 1 h, vacuum ltered, acetone removed (vacuum rotary evaporation, 35 C),
purged with nitrogen and stored in the dark at 20 C until analysis. Ethanol extraction was achieved using 70% ethanol adjusted to
pH 2.0 with formic acid for 15 h at ambient temperature followed
by solvent removal with vacuum ltration. The residue was re-extracted with hexane three times (1 h each), the supernatants were
pooled, vacuum ltered, and hexane removed as above to obtain
oil. Supercritical CO2 extraction was carried out based on AOAC
International method (2000) without co-solvent for 1 h in a laboratory-scale supercritical uid extraction system (Thar Technologies
Inc., Pittsburgh, PA) with carbon dioxide (99.95% purity, Praxair,
Edmonton, AB) compressed to 52 kPa with the vessel (500 ml)
temperature controlled at 100 C. Flaxseed hulls equilibrated to
23% moisture were hydraulically pressed (Carver Press, 280
kg/cm2) to extract cold-pressed oil. Commercial cold-pressed axseed oil (Omega Nutrition, Vancouver, BC) was used as control.
Extractions were performed in triplicate and analysed separately.
2.1. Analytical procedures
Thermal characteristics of oils were measured using a modulated differential scanning calorimeter (Modulated DSC-2910, TA
Instruments, New Castle, DE) described previously (Oomah, Dumon, Cardador-Martinez, & Godfrey, 2006). A ow of nitrogen
gas (100 ml/min) was used in the cell cooled by helium (150 ml/
min) in a refrigerated cooling system. The instrument was calibrated for temperature and heat ow with mercury (melting point,
mp = 38.8 C, TA Instruments standard), gallium (mp = 29.8 C, TA
Instruments standard) and indium (mp = 156.6 C, DH = 28.7 J/g,
Aldrich Chemical Co.). Oil samples (45 mg) were weighed in open
solid fat index (SFI) aluminium pans (T70529, TA Instruments). An
empty similar pan was used as reference. The sample and reference
pans were then placed inside the calorimeter and kept at 70 C for
5 min. The temperature was lowered from 70 to 65 C at a rate of
1 C/min with modulation at a period of 60 s and temperature
amplitude of 0.16 C. Samples were then kept at 65 C for
1 min, and then raised again at the same rate up to 70 C. Scans
were performed at 10 C/min. For thermal oxidation, pans were
cooled to 10 C and scanning was done by heating at 1 C/min to
350 C in the presence of oxygen (100 ml/min). Thermal oxidation
measurements were performed in duplicate.
Separation of individual lipid classes was performed using solid-phase extraction cartridge, (Bakerbond amino [NH2] disposable
extraction column, 500 mg, J.T. Baker Inc., Phillipsburg, NJ), with
aminopropyl packing, essentially as described by Oomah, Ladet,
Godfrey, Liang, and Girard (2000) based on the procedure of Carelli,
Brevedan, and Crapiste (1997). The cartridge was preconditioned
with 2 ml methanol, 2 ml chloroform, and 4 ml hexane before
use. A micropipette was used to inject 150 mg of oil dissolved in
chloroform. Lipid classes were recovered by sequential elution under vacuum (510 mm Hg) with 4 ml each of chloroform/isopropanol (2/1, v/v), diethyl ether/acetic acid (95/5, v/v), and methanol to
separate neutral lipids, free fatty acids, and phospholipids, respectively. The eluates were collected, evaporated under nitrogen,
weighed, and stored at 20 C. Acidic lipids were eluted with hexane/isopropanol/ethanol/0.1 M ammonium acetate/formic acid

(420/350/100/50/0.5) containing 5% phosphoric acid according to

Kim and Salem (1990), evaporated for 10 min and then extracted
with 1 ml of chloroform three times after adding 1 ml of water.
The chloroform fractions were combined, evaporated under nitrogen and weighed to determine yield.
2.2. Antioxidant assay
The photochemiluminescence (PCL) assay, based on the methodology of Popov and Lewin (1999) was used to measure the antioxidant activity of oils with a Photochem instrument (Analytik
Jena, USA Inc., Delaware, OH) against superoxide anion radicals
generated from luminol, a photosensitiser, when exposed to UV
light. Oil samples (100 mg) were diluted with hexane (1 ml) prior
to antioxidant determination described previously (Oomah, Tiger,
Olson, & Balasubramanian, 2006) using the ACL kit provided by
the manufacturer designed to measure the antioxidant activity of
lipophillic compounds. Antioxidant activity was monitored for
180 s and expressed as lm Trolox equivalent/g sample. Antioxidant assay was duplicated for each sample.
2.3. Extraction and analysis of secoisolariciresinol diglucoside
The lignan, secoisolariciresinol diglucoside (SDG) in defatted
axseed hulls was extracted and analysed essentially as described
previously (Ho, Cacace, & Mazza, 2007) by high performance liquid
chromatography based on the modied procedure of Muir and
Westcott (2000). Briey, defatted axseed hulls (0.5 g) were extracted by direct hydrolysis based on the protocol of Eliasson, Kamal-Eldin, Andersson, and man (2003) as described by Ho et al.
(2007). The hydrolysates were acidied (5 ml of 2 M H2SO4), centrifuged (11,000g, 10 min) and the supernatants (0.6 ml) were
mixed with methanol (0.9 ml, 30 min), re-centrifuged (11,000g,
5 min) and ltered (0.45 lm, Gelman Science Inc., Ann Arbor, MI)
prior to HPLC analysis. SDG was separated on a Luna C18, 5 lm,
100 , 250  3.00 mm column with a C18 Security Guard cartridge
(Phenomenex, Torrance, CA) using a Waters HPLC System (Waters
Corp., Milford, MA) consisting of a pump (Model 600), an autosampler (Model 717 plus), a degasser (Agilent 1100) and a 996 photodiode array detector operated by the Empower software. The linear
gradient elution with mobile phases consisting of 0.025% TFA in
water (solvent A) and methanol (solvent B) was carried out at
30 C at 0.4 ml/min for 70 min (Ho et al., 2007). SDG was detected
at 280 nm and quantied based on external SDG (ChromaDex,
Santa Ana, CA) calibration curves ranging from 10 to 400 lg/ml.
At least three determinations were made for all assays, except
for antioxidant activity which was determined in duplicate. Analysis of variance by the general linear models (GLM) procedure and
means comparisons by Duncans test were performed according
to Statistical Analysis System (SAS Institute, 1990).
3. Results and discussion
Oil yield from commercial axseed hulls (18.2%, Table 1) was
lower than the values (28%) reported earlier for hull fractions
of various cultivars obtained by dehulling (Oomah & Mazza,
1997; Oomah & Mazza, 1998). Fat content (18% and 27% dry matter) was also reported for axseed powder obtained by abrasion for
1 and 2 min, respectively (Myllymaki, 2002). Oil content of axseed hull obtained by the Urschel Comitrol Processor (Madhusudhan, Wiesenborn, Schwarz, Tostenson, & Gillespie, 2000) showed a
range (1517%) comparable to values observed in this study. The
highest yield was generally obtained from Natunola hulls (20.3%)
in accordance with the manufacturer fact sheet (minimum of
20% oil). Differences in extraction methods were statistically significant (P < 0.0002) due mostly to differences observed for Natunola

625

B.D. Oomah, L. Sitter / Food Chemistry 114 (2009) 623628

Table 1
Fractionation of axseed hull oils*.
Sample

Oil yield (%)

Neutral lipid

Free fatty acid

Phospholipids

Acidic lipids

Acetone
Cold press
Ethanol
Hexane
Petroleum Ether
Supercritical CO2
Omega
Natunola
Fortigrad
Mean

28.3a
9.10d
12.1cd
19.3b
18.3bc
20.4b
na
20.4x
15.7y
18.2

92.9a
93.6a
91.0ab
93.0a
87.8b
93.4a
92.8
93.1
91.8
92.5

2.09
1.66
2.01
2.34
2.56
2.30
1.37
1.73y
2.48x
2.05

2.33
3.24
2.36
2.25
4.35
3.26
3.68
2.94
3.03
3.05

2.70ab
1.45b
3.65ab
2.39ab
5.27a
1.01b
2.16
2.19
2.70
2.43

Means in a column followed by the same letter are not signicantly different by Duncans multiple range test at the 5% level. na, Not applicable.

hulls. Overall, extraction with acetone and cold press produced the
highest (28.2%) and lowest (9.1%) oil yield, respectively, independent of the source material. Variation in oil yield (18.320.4%)
(Table 1) obtained using hexane and petroleum ether and with
supercritical CO2 was not signicantly different. Hexane, ether
and supercritical CO2 are known to extract simple neutral lipids
whereas acetone also extracts polar lipids due to its high polarity
leading to increase in oil recovery. The supercritical CO2 method
yielded more oil (statistically not signicant at P = 0.05) than those
with petroleum ether or hexane contrary to lower (1034%) recoveries reported for axseed (Barthet & Daun, 2002; Bozan & Temelli,
2002). A comparative oil yield (20.5%) has been reported for rice
bran extracted with supercritical CO2 and hexane (Kuk & Dowd,
1998). Accelerated solvent extraction (ASE 100, Dionex Corporation, Sunnyvale, CA) with default method conditions (10 kPa,
105 C, 40 min) of Natunola axseed hulls produced oil
(22.9 0.08%, w/w) similar to that obtained by the lengthy petroleum extraction method (22.3 0.08%, w/w), thereby validating
the actual oil content of the material independent of methodology.
Flaxseed hull oils consisted primarily of neutral lipids (92.5%;
range 87.893.6%) with minor amounts of free fatty acids (2.1%;
range 1.42.6%), phospholipids (3.1%; range 2.254.35%) and acidic
lipids (2.4%; range 1.05.3%) of the crude oil (Table 1). Similar high
levels of neutral lipids (95.5%) and free fatty acids (0.31.6%) and
other lipid fractions (2.94.2%) have previously been reported for
axseed oil extracted from meal (Oomah et al., 1996). The range
of free fatty acids and phospholipids in axseed hull oils was lower
than those generally reported for rice bran oil (Dunford, 2005). Oil
extracted from both sources were similar in composition except
that average free fatty acid content of Fortigrad samples (2.48%)
was signicantly (P = 0.04) higher than those of Natunola (1.73%).
Petroleum ether extracted signicantly (P = 0.05) less neutral
lipids, almost twice the phospholipids and acidic lipids amount
(albeit not statistically signicant) than hexane despite their similar polarity. The differences in free fatty acids were not signicant
amongst extraction methods indicating similar ability in extracting
polar compounds. Supercritical CO2 and cold press extracted oils
were similar in composition since they had the highest and lowest
phospholipids and acidic lipid contents, respectively. The free fatty
acid content of the supercritical CO2 extracted oil from hulls was
twice the level reported for those obtained from axseed (Bozan
& Temelli, 2002). Neutral lipid content of axseed hulls showed
high negative (r = 0.72 and 0.86; P < 0.0001) correlation with
phospholipids and acidic lipids, respectively.
Flaxseed hull oil exhibited signals in both reversing and nonreversing components at approximately the same temperatures
(between 33 and 14 C) for the three thermal transitions indicating signal splitting. Two reversing transitions (between 35
and 33 C) and (between 25 and 24 C) indicative of crystalline melting were observed corresponding to the a and b polymorphic forms, respectively (Fig. 1). The second thermal transition

(24 to 25 C) reects the melting point of axseed oil (20 to

24 C) (Przybylski, 2005). A minor transition occurred at
14 C. The reversing component of the heat ow was highly sensitive to the initial source material (Table 2). For example, the rst
endothermic peak (the a-form) of Natunola oil occurred at signicantly higher temperature (32.3 C) suggesting increased stability than that of oil obtained from Fortigrad (34.9 C). Oil
extracted by cold press, acetone, and supercritical CO2 had higher
transition temperature than those obtained by other extraction
procedures. In the non-reversing component curves to which kinetic events such as crystallisation, crystal perfection and reorganisation are ascribed, three endotherms were observed in the 35 to
13 C region (Fig. 2). These endothermic transitions are indicative
of the crystallisation and recrystallisation of the metastable a form.
The rst endothermic peak (34 C with high activation energy of
25 to 19 J/g) suggests kinetic stability implying a rst-order
transition. The second and third peaks in the 25 to 13 C region
were assigned to b0 and b crystallisation forms, respectively, believed to be due to the transformation of the metastable phase
(a) to the more stable b form. Modulated DSC enabled the detection of small transitions in the reversing signal with great clarity.
These transitions representing apparent changes in heat capacity
in response to modulation were lower than endotherms observed
in non-reversing events. Thus, the endotherms of the rst and specically the second and third thermal reversing events were on
average only 69%, 30% and 39% of those of the non-reversing heat
ow, respectively (Table 2). Extraction procedures and source
material had no signicant effect on the DHa values for the reversing heat ow, although DHb values for both reversing and nonreversing heat ows of Natunola oils were signicantly
(P < 0.0001) higher than those of Fortigrad.
Flaxseed hull oil exhibited three maxima on the DSC oxidation
curves indicating that thermoxidation can be characterised by at

Heat Flow (W/g)

-0.02

-0.04

Hexane
Acetone
Ethanol
Petroleum ether
CO2
Cold Press

-0.06

-0.08
-60

-40

-20

20

Temperature (C)
Fig. 1. Modulated differential scanning colorimetry (MDSC) reversing component
of Fortigrad axseed hull oils.

626

B.D. Oomah, L. Sitter / Food Chemistry 114 (2009) 623628

Table 2
Thermal characteristics of axseed hull oils.
Sample

Acetone
Cold press
Ethanol
Goldsch
Hexane
Supercritical CO2
Omega
Natunola
Fortigrad
Overall Mean

Reversing heat ow

Non-reversing heat ow

Ta

D Ha

Tb

D Hb

Tc

D Hc

Ta

D Ha

Tb

D Hb

Tc

D Hc

31.8d
33.9d
33.4bc
33.4b
33.5bc
33.6c
33.1y
32.3x
34.9z
33.6

15.8
15.8
15.6
16.8
15.5
14.6
14.9
16.2
15.2
15.7

24.0
25.1
24.0
24.1
24.5
24.4
23.7x
23.6x
25.1y
24.4

2.2ab
2.6abc
2.2a
2.8c
2.7abc
2.2a
2.8y
3.2y
1.8x
2.5

15.1c
13.9b
13.8b
13.9b
13.3ab
14.9c
12.8x
13.3x
14.9y
14.1

0.2a
1.6b
2.3b
0.4a
0.3a
0.2a
3.8y
0.7x
1.1x
1.0

34.0cd
34.2d
33.9abcd
33.6a
33.8abc
34.0bcd
33.7y
32.5x
35.4z
33.9

23.2b
22.7b
19.4a
24.2b
23.5b
24.8b
18.0x
22.2y
23.6y
22.9

24.1ab
25.5c
24.0ab
24.0ab
25.0bc
24.4abc
23.6x
23.6x
25.4y
24.5

6.8ab
8.6c
7.9bc
8.7c
9.0c
8.2bc
6.4x
9.5y
6.9x
8.2

14.4c
14.3c
14.0bc
13.9bc
13.6b
13.9bc
13.0x
13.1x
14.9y
14.0

0.9a
4.5b
5.7b
1.3a
1.7a
1.1a
7.5y
1.8x
3.3x
2.6

300

350

Means in a column followed by the same letter are not signicantly different by Duncans multiple range tests at the 5% level.

4.5
3.5

Heat Flow (W/g)

Heat Flow (W/g)

0.01

-0.01
Hexane
Acetone
Ethanol
Petroleum ether
CO2
Cold Press

-0.03

-0.05

-0.07
-60

-40

-20

20

Temperature (C)

2.5

Hexane
Acetone
Ethanol
Petroleum ether
CO2
Cold Press

1.5
0.5
-0.5
50

100

150

200

250

Temperature (C)

Fig. 2. Modulated differential scanning colorimetry (MDSC) non-reversing component of Fortigrad axseed hull oils.

least three step exothermic effects (Fig. 3). These peaks could be
considered as an indication of the level of cross-linking. The oil
extracted using acetone, hexane, petroleum ether and supercritical CO2 showed an additional peak between 211 and 227 C indicating instability at this particular temperature. Oxidation of
axseed hull oil started at 105163 C, within the temperatures
reported for edible oils (130180 C) with mean onset and oxidation temperatures at 121 and 150 C, respectively and peaked at
194 to 200 C depending on extracting solvent and sample types
(Fig. 3; Table 3). The onset temperature (105130 C) corresponded to the ash point (120130 C) of axseed oil (Przybylski, 2005). The mean oxidation temperature of axseed hull oil
at 150 C resembled that of c-oryzanol in rice bran oil (Bucci,
Magri, Magri, & Marini, 2003). Oils extracted with petroleum
ether and ethanol had the lowest and highest onset, oxidation

Fig. 3. Differential scanning colorimety (DSC) of the thermo-oxidation proles of

Fortigrad axseed hull oils.

and peak1 temperatures, respectively. Signicant differences

(P < 0.001) were observed in onset, oxidation and peak1 temperatures amongst sample types (Omega, Natunola and Fortigrad),
with Fortigrad and Omega samples generally having the highest
and lowest temperatures, respectively. The third peak at 242 to
267 C differed signicantly (P < 0.0001) amongst the extracting
solvents; the oil with the lowest onset, oxidation and peak1 temperatures also had the highest peak3 temperature. The fourth
peak at 306320 C indicates the inability of oxygen uptake,
resulting in complete thermal polymerisation. Flaxseed hull oil
showed no signicant differences in the temperature of the fourth
peak; an observation similar to that reported previously for
hempseed and echinacea seed oils (Oomah, Busson, Godrey, &
Drover, 2002; Oomah et al., 2006). Comparison of Natunola with

Table 3
Thermoxidation temperatures (C) and antioxidant capacity of axseed hull oils*.
Sample

Onset

Oxidation temperature

Peak1

Peak2

Peak3

Peak4

Antioxidant capacity

SDG (mg/g)defatted hull

Acetone
Cold press
Ethanol
Petroleum ether
Hexane
Supercritical CO2
Omega
Natunola (N)
Fortigrad (F)
Mean (N and F)

123.8b
113.9c
130.0a
105.0d
121.2b
118.7bc
141.1x
128.3y
111.2z
120.9

154.3b
145.8c
162.5a
137.7d
146.4c
147.4c
164.1x
158.6y
140.9z
150.4

198.3ab
194.0c
200.3a
193.8c
196.8bc
197.3ab
203.2x
200.1y
193.9z
197.3

211.0b
nd
nd
225.5a
227.2a
210.9b
nd
225.5x
223.4y
224.3

248.8cd
242.2d
255.9bc
266.6a
259.8ab
251.2c
254.5xy
257.4x
249.0y
253.2

313.5ab
306.0b
318.1a
316.6a
320.3a
318.7a
322.0x
319.0xy
314.2y
317.1

0.94b
0.61c
0.93b
0.55c
0.59c
1.18a
1.13x
0.79y
0.89y
0.84

20.19d
45.93b
27.52c
51.72a
47.71b
53.08a
na
31.18y
48.49x
41.02

Means in a column followed by the same letter are not signicantly different by Duncans multiple range test at the 5% level. nd, Not detected. na, Not applicable.
Antioxidant capacity expressed as lm Trolox equivalent/g sample.

B.D. Oomah, L. Sitter / Food Chemistry 114 (2009) 623628

Fortigrad samples revealed that ethanol extract of Natunola and

cold press extract of Fortigrad had the highest and lowest onset,
oxidation, peaks1, 3 and 4 temperatures, respectively.
The antioxidant activity of axseed hull oil may be attributed to
its content of a-tocopherols since fractionated hull contains approximately 26% of the total tocopherols found in the whole seed (Oomah, Kenaschuk, & Mazza, 1997). Extraction methods had
signicant (P < 0.0001) effect on antioxidant activity of hull oils with
supercritical CO2 extracted oil exhibiting the highest antioxidant
activity, whilst those extracted with petroleum ether, hexane, or
cold pressed showed no signicant differences in antioxidant activity (Table 3). Antioxidant activity of Omega sample types was significantly (P < 0.0001) higher than those of Natunola and Fortigrad. The
comparatively high antioxidant activity of the supercritical CO2 extracted oil indicates the mild treatment and selectivity of carbon
dioxide in lipid solubilisation without affecting other components.
The cold-pressed oil exhibited low antioxidant activity, about half
that of the commercial (cold pressed, omega), in spite of its low free
fatty acid and acidic lipid content, suggesting that extracting conditions of the same procedure may alter the overall effectiveness of
antioxidant extraction. Decreasing the polarity of the solvent (particularly ethanol to hexane) lead to a signicant (P = 0.05) decrease
(0.900.60 lm/g) in oil antioxidant activity. Antioxidant activity of
the acetone extract was signicantly higher than those extracted
by hexane, a nding similar to those observed in wheat bran and
oat bran (Oufnac et al., 2007; Sun, Xu, Godber, & Prinyawiwatkul,
2006). A change in solvent polarity is known to alter its ability to dissolve selected group of antioxidant compounds thereby inuencing
the antioxidant activity estimation (Zhou & Yu, 2004). A case study
for recovering residual axseed oil by supercritical CO2 from the
after-press cake found the process to be technically and economically viable with extraction efciency greater than 95% (Martinez,
2005). Coincidentally, the supercritical CO2 extraction method
resulted in defatted axseed hull, the residual secondary product,
with the highest SDG content (53 mg/g) (Table 3). The SDG content
of axseed hull extracted by solvent systems decreased in the following order: CO2 = petroleum ether > hexane = cold press > ethanol > acetone. This order, except for CO2, is the exact opposite to
that observed with the antioxidant capacity indicating a negative
correlation (r = 0.556; p = 0.017) between SDG content of defatted
axseed hulls and antioxidant activity of the oil. Flaxseed hull defatted with petroleum ether and supercritical CO2 did not differ significantly (P < 0.0001) in SDG content contrary to reports by Zhang and
Xu (2007) where SDG yield by petroleum ether (7.6 g/kg) was lower
than those obtained by supercritical CO2 (10 g/kg). This could be due
to the differences in the SDG content of the starting material as evidenced in this study with Fortigrad hulls containing 55% higher SDG
than Natunola (Table 3). Thus supercritical CO2 extraction is an elegant strategy to obtain oil delivering both a-linolenic acid and
antioxidant benets of axseed hulls leaving behind a valuable
solvent-free secondary product with the highest SDG content.
Preprocessing of axseed hulls similar to those used for rice bran
(Orthoefer, 2005) may provide further improvement in superior
quality oil suitable for valuable nutraceutical products.
Acknowledgement
We thank David V. Godfrey, for his technical assistance.
References
AOAC International. (2000). Fat (crude) or ether extract in animal feed; Supercritical
uid extraction (SFE) method. AOAC Ofcial Method 920.39; 999.02. Ofcial
Methods of Analysis (17th ed.).
Barcel-Coblijn, G. (2007). a-Linolenic acid-enriched diets. A valid strategy to
increase n3 fatty acids levels. International News on Fats, Oils, and Related
Materials INFORM, 18(11), 719721.

627

Barre, D. E., Griscti, O., Mizier-Barre, K. A., & Hafez, K. (2005). Flaxseed oil and
lipoprotein (a) signicantly increase bleeding time in type 2 diabetes patients in
Cape Breton, Nova Scotia, Canada. Journal of Oleo Science, 54, 347354.
Barthet, V. J., & Daun, J. K. (2002). An evaluation of supercritical uid extraction as
an analytical tool to determine fat in canola, ax, solin, and mustard. Journal of
the American Oil Chemists Society, 79, 245251.
Bozan, B., & Temelli, F. (2002). Supercritical CO2 extraction of axseed. Journal of the
American Oil Chemists Society, 79, 231235.
Bucci, R., Magri, A. D., Magri, A. L., & Marini, F. (2003). Comparison of three
spectrophotometric methods for the determination of c-oryzanol in rice bran
oil. Annals of Bioanalytical Chemistry, 375, 12541259.
Carelli, A. A., Brevedan, M. I. V., & Crapiste, G. H. (1997). Quantitative determination
of phospholipids in sunower oil. Journal of the American Oil Chemists Society,
74, 511514.
Dunford, N. T. (2005). Germ oils from different sources. In F. Shahidi (Ed.), Baileys
industrial oil and fat products (6th ed., pp. 195231). Hoboken, NJ: John Wiley &
Sons, Inc.
Eliasson, C., Kamal-Eldin, A., Andersson, R., & man, P. (2003). High performance
liquid chromatographic analysis of secoisolariciresinol diglucoside and
hydrocinnamic acid glucosides in axseed by alkaline extraction. Journal of
Chromatography A, 1012, 151159.
Griel, A. E., Kris-Etherton, P. M., Hilpert, K. F., Zhao, G., West, S. G., & Corwin, R. L.
(2007). An increase in dietary n3 fatty acids decreases a marker of bone
resorption in humans. Nutrition Journal, 6, 2<http://www.nutritionj.com/
content/6/1/2>.
Ho, C. H. L., Cacace, J. E., & Mazza, G. (2007). Extraction of lignans, proteins and
carbohydrates from axseed meal with pressurized low polarity water.
Lebensmittel Wissenchaft und Technologie, 40, 16371647.
Kim, H-Y., & Salem, N. Jr., (1990). Separation of lipid classes by solid phase
extraction. Journal of Lipid Research, 31, 22852289.
Kuk, M. S., & Dowd, M. K. (1998). Supercritical CO2 extraction of rice bran. Journal of
the American Oil Chemists Society, 75, 623628.
Madhusudhan, B., Wiesenborn, D., Schwarz, J., Tostenson, K., & Gillespie, J. (2000).
A dry mechanical method for concentrating the lignan secoisolariciresinol
diglucoside in axseed. Lebensmittel Wissenchaft und Technologie, 33, 268
275.
Mantzioris, E., James, M. J., Gibson, R. A., & Cleland, L. G. (1994). Dietary substitution
with an a-linolenic acid-rich vegetable oil increases eicosapentaenoic acid
concentrations in tissues. American Journal of Clinical Nutrition, 59,
13041309.
Martinez, J. L. (2005). Recovery of residual oils after mechanical press using
supercritical uid extraction. International News on Fats, Oils, and Related
Materials INFORM, 16(10), 612613.
Muir, A. D., & Westcott, N. D. (2000). Quantication of the lignan secoisolariciresinol
diglucoside in baked goods containing ax seed or ax meal. Journal of
Agricultural and Food Chemistry, 48, 40484052.
Myllymaki, O. (2002). Processing of axseed. United States Patent, 6,440,479 B1.
Oomah, B. D. (2003). Processing of axseed ber, oil, protein, and lignan. In L. U.
Thompson & S. C. Cunnane (Eds.), Flaxseed for human nutrition (2nd ed.,
pp. 363386). Champaign, IL: AOCS Press.
Oomah, B. D., Busson, M., Godfrey, D. V., & Drover, J. C. G. (2002). Characteristics of
hemp (Cannabis sativa L.) seed oil. Food Chemistry, 76, 3343.
Oomah, B. D., Dumon, D., Cardador-Martinez, A., & Godfrey, D. V. (2006).
Characteristics of Echinacea seed oil. Food Chemistry, 96, 304312.
Oomah, B. D., Kenaschuk, E. O., & Mazza, G. (1997). Tocopherols in axseed. Journal
of Agricultural and Food Chemistry, 45, 20762080.
Oomah, B. D., Ladet, S., Godfrey, D. V., Liang, J., & Girard, B. (2000). Characteristics of
raspberry (Rubus idaeus L.) seed oil. Food Chemistry, 69, 187193.
Oomah, B. D., & Mazza, G. (1997). Effect of dehulling on chemical composition and
physical properties of axseed. Lebensmittel Wissenchaft und Technologie, 30,
135140.
Oomah, B. D., & Mazza, G. (1998). Fractionation of axseed with a batch dehuller.
Industrial Crops and Products, 9, 1927.
Oomah, B. D., Mazza, G., & Przybylski, R. (1996). Comparison of axseed meal lipids
extracted with different solvents. Lebensmittel Wissenchaft und Technologie, 29,
654658.
Oomah, B. D., Tiger, N., Olson, N., & Balasubramanian, P. (2006). Phenolics and
antioxidative activities in narrow-leafed lupins (Lupinus angustifolius L.). Plant
Foods for Human Nutrition, 61, 9197.
Orthoefer, F. T. (2005). Rice bran oil. In Baileys industrial oil and fat products (6th ed.,
pp. 465489). Hoboken, NJ: John Wiley & Sons, Inc.
Oufnac, D. S., Xu, Z., Sun, T., Sabliov, C., Prinyawiwatkul, W., & Godber, J. S. (2007).
Extraction of antioxidants from wheat bran using conventional solvent and
microwave-assisted methods. Cereal Chemistry, 84, 125129.
Paschos, G. K., Magkos, F., Panagiotakos, D. B., Volteas, V., & Zampelas, A. (2007).
Dietary supplementation with axseed oil lowers blood pressure in
dyslipidaemic patients. European Journal of Clinical Nutrition, 61, 12011206.
Popov, I., & Lewin, G. (1999). Antioxidative homeostasis: Characterization by
means of chemiluminescent technique. In L. Packer (Ed.), Methods in
enzymology, Volume 300, Oxidants and Antioxidants Part B (pp. 96100).
Academic Press.
Przybylski, R. (2005). Flax oil and high linolenic oils. In F. Shahidi (Ed.), Baileys
industrial oil and fat products (6th ed., pp. 281301). Hoboken, NJ: John Wiley &
Sons, Inc.
SAS Institute Inc. (1990). SAS/STAT users guide, version 6 (4th ed.). Cary, NC: SAS
Institute.

628

B.D. Oomah, L. Sitter / Food Chemistry 114 (2009) 623628

Stark, A. H., Crawford, M. A., & Reifen, R. (2008). Update on alpha-linolenic acid.
Nutrition Reviews, 66, 326332.
Sun, T., Xu, Z., Godber, J. S., & Prinyawiwatkul, W. (2006). Capabilities of oat extracts
in inhibiting cholesterol and long chain fatty acid oxidation during heating.
Cereal Chemistry, 83, 451454.

Zhang, W., & Xu, S. (2007). Microwave-assisted extraction of secoisolariciresinol

diglucoside from axseed hull. Journal of the Science of Food and Agriculture, 87,
14551462.
Zhou, K., & Yu, L. (2004). Effects of extraction solvent on wheat bran antioxidant
activity estimation. Lebensmittel Wissenchaft und Technologie, 37, 717721.