BERRY SEEDS: A SOURCE OF SPECIALTY OILS WITH HIGH

CONTENT OF BIOACTIVES AND NUTRITIONAL VALUE

V. VAN HOED1,5, N. DE CLERCQ2, C. ECHIM1, M. ANDJELKOVIC1,3,
E. LEBER4, K. DEWETTINCK2 and R. VERH1
1

Department of Organic Chemistry

2
Department of Food Safety and Food Quality
Laboratory of Food Technology and Engineering
3

Research Group, Food Chemistry and Human Nutrition, Department of

Food Safety and Food Quality
Faculty of Bioscience Engineering
Ghent University
Coupure Links 653, 9000 Ghent
4

Aprs-Vin Enterprises, Inc. Post Office Box 2, Prosser, Washington

Submitted for Publication August 14, 2008
Revised Received and Accepted November 27, 2008

ABSTRACT
Selected berry seed oils from blackberry, blueberry, cranberry, strawberry, red raspberry and kiwi were characterized for their quality and nutritional characteristics. These oils are by-products of berry juice production
that have only recently gained commercial interest. Free fatty acid content was
below 1.6% for all examined oil samples. Peroxide value ranged between 0.6
and 44 mg O2/kg oil for blackberry and kiwi seed oils, respectively, and
p-anisidine value varied from 6 in cranberry to 23 in strawberry. Linolenic
acid content ranged from 17.53% in blackberry seed oil to 57.60% in kiwi seed
oil. The oxidative stability of all oils was rather low (0.17 h for kiwi to 8.4 h for
blackberry at 97.8C). Phytosterol contents ranged between 403 and 692 mg/
100 g for blackberry and cranberry, respectively. The content of tocols
(tocopherol + tocotrienol) varied from 34.4 for kiwi to 2,133 mg/kg for red
raspberry seed oils.

Corresponding author. TEL: 32-9-264-58-59; 32-9-264-59-50 (secretariat); FAX: 32-9-264-62-43;

EMAIL: vera.vanhoed@ugent.be

Journal of Food Lipids 16 (2009) 3349. All Rights Reserved.

2009, Wiley Periodicals, Inc.

33

34

V. VAN HOED ET AL.

PRACTICAL APPLICATIONS
A waste stream of fruit processing is used to extract the oil from berry
seeds. Such oils are particularly rich in essential fatty acids (with a favorable
low n-6/n-3 ratio) and antioxidants. They are incorporated in cosmetic preparations such as hand and body creams, and shampoos. Their composition is
also interesting from a nutritional point of view. As the commercial interest is
growing, chemical studies are necessary to elucidate the composition, activity
and stability of different berry seed oils.

INTRODUCTION
The healthful effects of minor components present in vegetable oils have
become a subject of renewed interest in recent years (Shahidi 2000). Many
studies have been conducted on the valuable constituents in extra virgin olive
oil and other cold pressed bulk oils. Nowadays, specialty high-value vegetable
oils (like berry seed oils) are gaining attention owing to their health benefits
which are linked to their high content of polyunsaturated fatty acids (PUFAs)
and antioxidants.
Production of value-added products out of waste streams from agricultural products may also offer a possible environmental and economical benefit.
In the production of wines from grapes and juices from berries, the liquid is
separated from a waste stream, which is a paste containing the peel and seeds.
The seeds can be separated from the paste and a value-added product, seed oil,
can be obtained by cold-pressing, leaving the seed-flour as a side stream. Both
product streams are gaining more and more attention, constituting a small but
growing market segment.
The amount of caneberry seeds (Rubus spp.) from Oregon and Washington seedless berry production in 2003 was estimated to be 180,000 kg (Luther
et al. 2007). In Canada, 14.4 thousand tons of raspberries were produced in
2005, containing about 10% of seeds with an oil content of 23%. These values
are only a fraction of those emerging during bulk oil production. However, in
the case of berry seed oils, owing to the cold and chemical-free processing,
valuable compounds are preserved and it is possible to isolate them. For
example, cranberry press cake (Zheng and Shetty 2000) and grape pomace
(Baydar et al. 2007; Bail et al. 2008) have been used for the preparation of
food grade antioxidant extracts.
Other reported sources for berry seed oils are red raspberry, black raspberry, boysenberry, marion blackberry, evergreen blackberry, blueberry, strawberry and cranberry (Oomah et al. 2000; Wang and Lin 2000; Parker et al.
2003; Bushman et al. 2004; Parry and Yu 2004; Parry et al. 2005).

BIOACTIVES OF BERRY SEEDS

35

All berry seed oils have a high content of PUFAs in common, providing
essential fatty acids (EFAs). These are fatty acids that cannot be synthesized
within an organism from other components by any known chemical pathway;
and therefore must be obtained from the diet. The higher consumption level
of n-6 fatty acids compared with n-3 fatty acids is perceived to be a concern
when considering the prevention of cancers, heart diseases, hypertension and
autoimmune disorders (Connor 2000; Hung et al. 2000; Aronson et al. 2001).
In the Western diet the present n-6/n-3 ratio has been estimated to be between
10 and 25 to 1 (Parry et al. 2005; Bawa 2008), while the recommended ratio
is estimated to be 4 to 1 (Parry et al. 2005). The n-6 to n-3 PUFA imbalance
has been brought about primarily by the increase in vegetable and seed oil use,
and a decreased intake of fish, fish oil and vegetable oils containing a high
level of n-3 fatty acids. Berry seed oils on the other hand have a favorable
n-6/n-3 FA ratio compared with some other vegetable oils (Parker et al. 2003;
Parry et al. 2006). Moreover, these seed oils are also rich in various antioxidants, which are related to a protective effect against cardiovascular lipid
oxidation and antitumor activities (Wang and Jiao 2000).
In the case of grape seeds, oils and flours are known to contain high
contents in EFAs, vitamin E and polyphenols (Baydar et al. 2007), being
correlated with the high oxygen radical absorption capacity values (Wettasinghe and Shahidi 1999). Those products are already commercialized in the
U.S.A. (Leber and Ramonas 2008).
Recently, the properties of some berry seed oils have been reported in
the literature. Parry and Yu (2004) and Parry et al. (2005) found significant
amounts of a-linolenic acid (19.634.2 g per 100 g of oil) and tocopherols
(2602,277 mmoles/kg oil), polyphenols (0.092.00 mg gallic acid equivalents per gram of oil) and carotenoids (12.530.0 mmoles/kg oil) in marionberry, boysenberry, red raspberry and blueberry seed oils. Furthermore,
the oxidative stability of the oils (OSI) and the capacity of the antioxidant
extracts to scavenge oxygen, 2,2-azinobis-3-ethylbenzothiazoline-6-sulfonic
acid (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were
studied.
All these data indicate the high nutritional value of berry seeds and
derivatives. These properties could justify the commercialization of the oils for
health-conscious consumers at high prices. However, as is well known for
extra-virgin olive oil, the high price can induce fraudulent mixing of highvalue oils with lower-value bulk oils. Hence, both from a nutritional point of
view, and for authentication purposes, the characterization of the chemical
composition of oils remains an important issue. In this respect, the chemical
composition is well known for the most common vegetable oils, especially
olive oil. Nevertheless, information is lacking concerning these compounds in
berry seed oils.

36

V. VAN HOED ET AL.

In the current work, cold-pressed and filtered oils from blackberry, red
raspberry, cranberry, strawberry, blueberry and kiwi seeds are studied. Basic
quality parameters such as free fatty acid content, peroxide value (PV) and
p-anisidine value (AnV) were determined. In addition, compounds with high
value in the oils potential as functional foods were determined such as the
fatty acid composition, including EFAs, unsaponifiable matter and tocols
(tocopherol + tocotrienol) fraction. The possibility to use novel oils in food
products is also dependent on their shelf life and necessary storage conditions
for maintaining their healthy nature. The oxidative stability index was compared with common vegetable oils (corn oil, soybean oil) and correlated with
the chemical composition of the oils tested.

MATERIALS AND METHODS

Berry Seed Oils
The berry seed oils were provided by Aprs-Vin Enterprises (Prosser,
WA). The berry fruits were all grown in the State of Washington in the Pacific
Northwest region of the U.S.A., between the towns of Everett and Bellingham.
Kiwi fruits were grown in farms in Californias Central Valley between the
cities of Sacramento and Bakersfield, U.S.A. Oils from six different berries,
namely red raspberry (Rubus idaeus) (RROL), blackberry (Rubus fruticosus)
(BBOL), cranberry (Vaccinium macrocarpon) (CBOL), blueberry (Vaccinium
corymbosum) (BUOL), strawberry (Fragaria F. x ananassa) (STOL) and kiwi
(Actinidia chinenesis) (KWOL), were studied.
Within 24 h of pressing the berries to make juice, the pomace was collected by FruitSmart (Prosser, WA) and dried within 10 h of collection (from
about 80% to 8% moisture). The seeds were mechanically separated (approx.
99.9% pure seed), and were double wrapped in heavy-duty polyethylene bags
and stored in dry-ambient-temperature cardboard boxes, before being pressed.
The oils were cold-pressed in a Tby Type 40a expeller press (rebro,
Sweden) at temperatures ranging from 40.6C to about 60.0C, depending on the
seed type and quality. Finally, the oils were cold-filtered at a temperature
below 10C and the raw oils were drawn through tubes immersed in ice-water
before passing through a plate filter Model MFP320-10 (Parkson, Fort Lauderdale, FL) with 11.5% of diatomaceous earth (grades FW 60 and FP 4,
Eagle Picher, Quincy, WA) for polishing (at or below 10C).
Free Fatty Acids
The amount of free fatty acids was determined by dissolving a known
quantity of fat in an ether/ethanol (50:50, v/v) mixture and by titration with

BIOACTIVES OF BERRY SEEDS

37

aqueous potassium hydroxide (0.1 M KOH) with phenolphthalein as an indicator. The acidity is a conventional expression for the percentage of free fatty
acids, expressed as g oleic acid per 100 g of sample. Analyses were carried out
in duplicate.
Fatty Acid Profile, Determined as Fatty Acid Methyl Esters
The fatty acid profile was determined by gas chromatography (GC) after
saponification of the triacylglycerols and methylation of the fatty acids. Fatty
acid methyl esters (FAMEs) were then analyzed using an Interscience Thermofocus GC (Interscience, Louvain-la-Neuve, Belgium) with a flame ionization detector (FID) and an autosampler. An RTX-2,330 capillary column
(cyanopropyl polysiloxane, 60 m 0.25 mm 2 mm, Interscience, Louvainla-Neuve, Belgium) was used with hydrogen as the carrier gas at a flow rate of
1 mL/min. One microliter of sample solution was injected via a split/splitless
injector (split ratio 1.60) at an inlet temperature of 230C. The oven temperature was 120C for 4 min, increased from 120C to 210C at 2C/min and held at
210C for 25 min. Detection was performed with the FID set to 250C. The
separated FAMEs were identified by retention time and quantified by determining the relative peak area. Single determinations were sufficient because
the method has a coefficient of variation below 0.03%.
Peroxide Value
The acetic acid isooctane method was used to determine the PV
(AOCS 1996a). This method determines all substances, in terms of milliequivalents of oxygen per 1,000 g of sample that oxidize potassium iodide
under the conditions of the test. These substances are generally assumed to be
peroxides or other similar products of fat oxidation. Analyses were carried
out in triplicate.
p-Anisidine Value
The AnV (unitless) defines the amount of aldehydes in a lipid sample
by reaction in an acetic acid solution of the aldehydic compounds present in
the samples with p-anisidine. The absorbance is read at 350 nm (AOCS
1996b). Analyses were carried out in duplicate.
Oxidative Stability Index
The oxidative stability index was determined following the AOCS (1997)
Official Method Cd 12b-92. Analyses were carried out in duplicate. In this
method the oil is subjected to oxidative conditions at 97.8C. The oil is heated
at a high temperature while blowing air through the sample (5 g), which is

38

V. VAN HOED ET AL.

captured in a tube with distilled water (50 mL). As the oxidation proceeds, the
air will contain more oxidation products, which increase the conductivity of
the water. The time at which the 1st derivative of the conductivity curve
increases is called the OSI time and represents the time taken to reach a
maximum level of conductivity (expressed in hours). High OSI values are
associated with a longer shelf life of the oil.
Unsaponifiable Matter
To determine the unsaponifiable matter, 2 mL of internal standard solution (0.1% of dihydrocholesterol in chloroform) was added to 1.5 g of oil
sample. After evaporation of the chloroform, the sample was saponified with
6 mL of a 6 M solution of KOH in water and 10 mL of ethanol. The solution
was refluxed for 90 min at 90C. After saponification, 10 mL of water was
added and the unsaponifiables were extracted first with 15 mL of petroleum
ether and then with 15 mL of diethyl ether. The combined extracts were
washed twice with 7 mL of a 0.5 M KOH solution in water and with 7 mL of
a 5% NaCl solution in water until the pH of the washing water was neutral. The
organic fraction was dried over MgSO4. After filtration and evaporation of the
solvent, the unsaponifiables were derivatized at 80C for 30 min after adding
0.5 mL of pyridin and 1.0 mL of (BSTFA + 1% TMCS) as silylating agent.
The derivatized samples were analyzed within 6 h after silylation.
Gas chromatographic analysis was performed on an Agilent 6,890 Series
GC System (Palo Alto, CA) equipped with an EC5 capillary column
30 m 0.25 mm 0.25 mm (Alltech, Deerfield, IL) and an FID. The velocity
of the carrier gas (helium) was 20 cm/s. The injection volume was 0.5 mL, with
a split ratio of 1:10. The injector and detector temperatures were 310C and
360C, respectively. The initial oven temperature was 285C for 30 min,
increased from 285C to 310C at 10C/min and held there for 10 min.
The identification was performed by comparing the relative retention
times and the mass spectra (obtained by GC-MS) with those of known standards and with literature. Phytosterols were quantified using the response
factor Rf = 1 relative to the internal standard dihydrocholesterol, while
squalene was quantified using the Rf = 1.07 (determined using pure dihydrocholesterol and squalene standards).
Tocopherols and Tocotrienols (Tocols)
Tocopherol and tocotrienol content was analyzed by normal phase HPLC
(Alltima Silica 5U, 250 mm 4.6 mm i.d., particle size 5 mm; Alltech,
Lokeren, Belgium) based on the AOCS (1996c) Official Method Ce 889.
Analyses were carried out in triplicate.

BIOACTIVES OF BERRY SEEDS

39

RESULTS AND DISCUSSION

Quality Parameters Acidity, Peroxide Value and p-Anisidine Value
Free fatty acids (%FFA) in oils represent a first quality parameter of the
samples. All oils had a %FFA between 0.49 (RROL) and 1.54 (STOL)%,
expressed as oleic acid equivalents (Table 1). As there are no guidelines
yet for these relatively new oils, the guidelines for olive oil were taken as a
reference. According to EC Regulation No 2,568/91 (EC Regulation of the
European Commission 1991) an olive oil belongs to the quality category of
extra virgin if %FFA < 0.8%; virgin if %FFA < 2%; and ordinary when
%FFA < 3.3%. Thus, from the measured values it can be concluded that the
seed oils correspond to the extra virgin and virgin categories.
To appraise the quality of oil samples it is also important to investigate the
oxidative status because the oxidative processes in oils and fats are one of the
main causes of the deterioration of the principal organoleptic and nutritional
characteristics of foodstuffs. The complex oxidation process can be summarized into two phases: first fatty acids react with oxygen to produce peroxides
as odorless compounds, which are then degraded into volatile aldehydes and
other compounds, responsible for the rancid odor and flavor, and also a
non-volatile fraction. The primary oxidation products are measured as PV and
the secondary products as AnV. The PV indicates the actual oxidative status
and the p-anisidine value indicates its history from the oxidative point of view.
To combine both, a TOTOX number can be calculated: TOTOX = AnV
+ (2 PV).
As presented in Table 1 it is clear that all the berry seed oil samples
were highly susceptible to oxidation. This is due to the high degree of unsaturation of all the oils examined. The reference values to ensure the oxidative
stability of a highly unsaturated oil are 5 meq O2/kg oil for the PV value and
a value of 20 (unitless) for the AnV (Health Canada, 2007). The PV is
significantly lower for BBOL and CBOL compared with the other oils and
they also show low AnV. The TOTOX value for these two oils is lower than
the maximum value (30), calculated from the maximum PV and AnV values,
and therefore acceptable. All the other oils had a TOTOX of 60 and more.
Also, for BBOL to STOL a higher AnV value was observed together with
higher FFA content (indicative for hydrolytic reactions occurring during prolonged storage).
Fatty Acid Methyl Esters
All seed oil samples contained high amounts of unsaturated fatty acids
ranging from 90.51% (KWOL) to 96.18% (RROL) (Table 2). RROL contained the highest amount of PUFAs (85.16%) and CBOL the highest quantity

0.59 0.22
24.62 1.74
8.41 0.47
44.37 0.22
42.88 0.28
26.25 0.32

0.96 0.00
0.67 0.00
0.70 0.05
1.20 0.00
0.49 0.01
1.54 0.04

BBOL
BUOL
CBOL
KWOL
RROL
STOL

10.80 0.49
10.30 2.45
6.18 1.06
22.41 5.04
16.00 0.23
22.74 1.54

AnV
11.97 0.65
59.54 4.23
29.06 1.64
111.16 5.06
101.76 0.61
75.23 1.67

TOTOX
8.23 0.81
2.55 0.71
6.33 1.10
0.18 0.04
3.43 0.39
4.28 0.39

OSI at 97.8C
(h)

27.7
8.5
21.3
0.4
11.5
14.3

PREDICTED OSI
at 80.0C (h)

The OSI value at 80C was estimated using the OSI instruments software, with the relationship OSI (97.8C) = 0.2949 OSI (80C) + 0.0517.
FFAs = free fatty acids; PV = peroxide value; AnV = p-anisidine value; TOTOX = total oxidation value = AnV + 2 PV; OSI = oxidative stability index;
BBOL = blackberry seed oil; BUOL = blueberry seed oil; CBOL = cranberry seed oil; KWOL = kiwi seed oil; RROL = red raspberry seed oil;
STOL = strawberry seed oil.

PV
(mg O2/kg oil)

FFAs (%)

Sample

TABLE 1.
QUALITY PARAMETERS FREE FATTY ACIDS, PEROXIDE, P-ANISIDINE, TOTOX, AND OSI VALUE OF THE STUDIED BERRY SEED OILS

40
V. VAN HOED ET AL.

BIOACTIVES OF BERRY SEEDS

41

TABLE 2.
FATTY ACID COMPOSITION OF THE COLD PRESSED SEED OILS (ABBREVIATION
LETTER BETWEEN BRACKETS)
Fatty acid

C12:0 (La)
C14:0 (M)
C16:0 (P)
C18:0 (S)
C18:1c (O)
C18:2n-6 (L)
C18:3n-3 (Ln)
C20:0 (Ar)
SAFAs
MUFAs
PUFAs
n-6/n-3

Fatty acid composition g/100 g fatty acids

BBOL

BUOL

CBOL

KWOL

RROL

STOL

0.04
0.05
3.71
2.18
14.72
61.22
17.60
0.47
6.45
14.81
78.74
3.48

0.02
0.09
5.66
1.78
21.42
42.51
28.28
0.25
7.80
21.43
70.77
1.50

0.14
0.08
5.38
1.25
25.30
37.68
30.09
0.07
6.88
25.14
67.98
1.25

0.02
0.03
5.96
3.09
14.60
17.55
58.40
0.34
9.39
14.60
76.01
0.31

0.07
2.43
0.90
10.87
53.67
31.68
0.37
3.82
11.02
85.16
1.69

0.05
4.32
1.68
14.55
42.22
36.48
0.71
6.85
14.71
78.44
1.16

BBOL = blackberry seed oil; BUOL = blueberry seed oil; CBOL = cranberry seed oil; KWOL = kiwi
seed oil; RROL = red raspberry seed oil; STOL = strawberry seed oil; SAFAs = saturated fatty acids;
MUFAs = monounsaturated fatty acids; PUFAs = polyunsaturated fatty acids; La = lauric acid;
M = myristic acid; P = palmitic acid; S = stearic acid; O = oleic acid; L = linoleic acid; Ln = linolenic
acid; Ar = arachidic acid.

of monounsaturated fatty acids (MUFAs: 25.14%) compared with the other

seed oils. All seed oils were a good source of EFAs. In all oils studied, except
KWOL, C18:2 was the most prevalent fatty acid, ranging from 60.98% in
BBOL to 37.44% in CBOL. KWOL merely contained 17.31% C18:2 but it had
a very high proportion of C18:3 (57.06%) which is reflected in a very low ratio
of n-6/n-3 (0.31). BBOL on the contrary, exhibited a high ratio of n-6/n-3
(3.58). All the oils exhibited a very favorable n-6/n-3 ratio, compared with
other vegetable oils. Surprisingly, the KWOL also contained a significantly
higher percentage of saturated fatty acids of 9.38%. When the seed oils within
the same genus e.g. Rubus gene (BBOL and RROL) are compared, differences
were observed for all fatty acids. On the other hand, in the oils of the Vaccinium genus (BUOL and CBOL) only the amounts of C18:2 and C18:3 were
different.
From the fatty acid profile it is clear that berry seed oils are very interesting from a nutritional point of view. These results are in line with those
reported in the literature (Bushman et al. 2004; Nawar 2004; Parry et al. 2005;
Parry et al. 2006; Oh et al. 2007). It is also important to notice that chemical
characterization of KWOL has not yet been reported. Similarly for STOL, only
limited literature data are available.

42

V. VAN HOED ET AL.

Unsaponifiable Matter
New data show that some clinically important properties of dietary fats
may be due to the presence of minor components such as phytosterols,
squalene or oxysterols, rather than to the triacylglycerol fatty acids alone
(Ostlund et al. 2002). Phytosterols effectively reduce LDL-cholesterol when
given as supplements, and the smaller amounts present in natural foods also
appear to be important (Ostlund 2004).
Both desmethylsterols and squalene were quantified in the unsaponifiable
fraction of the berry seed oils. In Table 3 the phytosterol content in the samples
is presented. Only the desmethylsterols were quantified, whereas a- and bamyrin, and cycloartenol were detected, but not quantified owing to co-elution.
The total desmethylsterol content varied widely between the different
berry seed oils, ranging from 404 mg/100 g for BBOL to 692 mg/100 g for
CBOL. To the best of our knowledge, literature data are only available for the
last one. Nawar (2004) found 657 mg/100 g of phytosterols in cranberry seed
oil, which is comparable to the results obtained in this study. The two highest
values were observed for the two oils of the Vaccinium genus (CBOL, BUOL).
Compared with known vegetable oils, all these values are higher than the sterol
content of olive oil (102206 mg/100 g) (Salvador et al. 2001), slightly higher
than that in soybean oil (184409 mg/100 g) or sunflower seed oil (244
455 mg/100 g), but (much) lower than the amount in corn oil (7952,215 mg/
100 g), or rice bran oil (1,055 mg/100 g) (Firestone 2006).
As in most vegetable oils, the main sterol in all samples was b-sitosterol,
ranging from 59.1% to 84.7% of the desmethylsterols. In the CBOL, the other
phytosterols were D7-sitosterol, constituting almost one-third of the total sterol
fraction, campesterol, D7-avenasterol and stigmasterol. This is in accordance
with the values reported by Nawar (2004). The phytosterol profile seems to be
relatively the same for fruits of the same species, even when produced under
different growing conditions. Thus, this could be a useful parameter in authentication studies.
The sterol composition of BUOL from the Vaccinium genus differed most
markedly from the CBOL in the content of D7-sitosterol, characteristic for
CBOL, and D5-avenasterol was detected only in BUOL. In BBOL and RROL,
both species of the Rubus genus, 403.7 and 493.7 mg phytosterols
per 100 g were found, respectively, consisting of very similar contents of
b-sitosterol, D5-avenasterol, campesterol, D7-avenasterol and stigmasterol.
The major difference between both Rubus oils was that in RROL 5.7%
D7-sitosterol was found, while in BBOL D7-sitosterol was only detected only
in trace amounts.
The KWOL was characterized by the lowest b-sitosterol content of all
oils analyzed, and very similar contents for D5-avenasterol and D7-avenasterol

580.2 48.6
3.4 0.0
0.3 0.0
66.5 0.1
13.8 0.7
10.7 0.1
5.3 1
84.0 0.7
16.0 0.2
178.1 21.2

403.7 3.8

5.3 0.1
1.8 0.0
84.7 0.2
7.0 0.2
tr
1.3 0.0
98.7 2.6
1.3
17.0 0.1

4.2 0.1
1.3 0.1
60.4 0.3
tr
31.6 0.2
2.5 0.0
65.9 2.1
34.1 2.5
671.5 16.7

692.3 3.65

CBOL

2.1 0.2
2.4 0.0
59.1 0.6
13.1 0.0
9.6 0.1
13.8 0.4
76.6 0.6
23.4 0.6
826.2 17.0

422.1 16.3

KWOL

4.5 0.0
1.2 0.0
79.6 0.1
7.2 0.2
5.7 0.2
1.8 0.0
92.5 1.9
7.5 1.2
8.4 0.4

493.7 2.2

RROL

5.4 0.3
2.3 0.1
71.1 0.7
8.7 0.1
8.2 0.1
4.3 0.1
87.5 0.8
12.5 0.4
tr

489.4 29.8

STOL

D5-avenasterol co-eluted with sitostanol for BUOL, KWOL, STOL, and with (a trace of) b-amyrin for RROL, CBOL and BBOL.
D7-sitosterol co-eluted with a trace of cycloartanol for CBOL, and was detected as a trace impurity in the cycloartanol peak of BBOL.
tr = trace, detected but not quantified. BBOL = blackberry seed oil; BUOL = blueberry seed oil; CBOL = cranberry seed oil; KWOL = kiwi seed oil;
RROL = red raspberry seed oil; STOL = strawberry seed oil.

Total desmethylsterols (mg/100g)

Sterols in %
Campesterol (D5)
Stigmasterol (D5)
b-sitosterol (D5)
D5-avenasterol
D7-sitosterol
D7-avenasterol
Sum D5 sterols (%)
Sum D7 sterols (%)
Squalene (mg/100 g)

BUOL

BBOL

TABLE 3.
STEROL CONTENT AND COMPOSITION OF SEED OILS

BIOACTIVES OF BERRY SEEDS

43

44

V. VAN HOED ET AL.

on one hand, and for campesterol and stigmasterol on the other hand,
with a moderate amount of D7-sitosterol. The high D7-avenasterol content is
characteristic for the KWOL. The last oil, STOL, did not show any characteristic sterol, but intermediate values for total sterols, and for the D5- and
D7-sterols.
The content of squalene, which is a precursor of phytosterols, and also an
antioxidant (Khallouki et al. 2003) varied widely among samples. A very high
value (671.5 mg/100 g) was found for CBOL which had the highest sterol
content but the highest squalene content was noted for KWOL (826.2 mg/
100 g), which contained a moderate amount of phytosterols. Compared with
other vegetable oils, CBOL and KWOL approached the highest levels of
squalene found typically in olive oil (8001,200 mg/100 g; Grigoriadou et al.
2007, or 599.0 mg/100 g; Tuberoso et al. (2007). Other seed oils with high
squalene contents are argan oil with 320 mg/100 g (Khallouki et al. 2003) and
pumpkin seed oil with 353 mg/100 g (Tuberoso et al. 2007). Very low values
were found in the two Rubus species (BBOL: 17.0 and RROL: 8.4 mg/100 g),
and for STOL (5.8 mg/100 g). The latter values were comparable to sunflower
seed oil (17.1 mg/100 g), rapeseed oil (43.7 mg/100 g) and corn oil (33.9 mg/
100 g); soybean oil and grape seed oil are devoid of squalene according to
Tuberoso et al. (2007).
Tocopherols and Tocotrienols (Tocols)
The highest tocopherol content was found in RROL (2,113 mg/kg), followed by CBOL (1,532.7 mg/kg) and BBOL (1,388.7 mg/kg), while all other
oils contained much lower amounts (Table 4). The lowest value was found for
KWOL (only 34.4 mg/kg). The total tocopherol (T) and tocotrienol (T3)
content may depend on different growing, processing and storage conditions,
and consequently may vary considerably within a certain oil class (Boskou
2006). For corn oil, 3313,716 mg/kg tocopherols (T) and 0709 mg/kg tocotrienols (T3) were reported. Soybean, on the other hand, contained 573
3,363 mg/kg T and 0172 mg/kg T3, mainly g-tocopherol. Olive oil contains
only 70150 mg/kg T (Firestone 2006).
All oils in this study contained g-tocopherol, which was by far the
main tocol in the Rubus species (BBOL, RROL: 1,311.7 and 1,640.7 mg/kg,
respectively). CBOL showed a unique composition with 1,235.0 mg/kg of
g-tocotrienol, and 152.7 mg/kg of a-tocotrienol. It has been reported that
tocotrienols exhibit a higher biopotency than tocopherols against different
diseases (Nakagawa et al. 2007). Except for palm, wheat germ and rice bran
oil, tocotrienols are not very common in other vegetable oils (Firestone 2006).
This means that CBOL represents a valuable alternative source of tocotrienols.
b-tocopherol and b-tocotrienol were not detected in any of the samples tested.

4.4 0.2

34.4 0.1
330.4 11.4

6.0 1.0
375.2 12.4

25.4 6.5

1,311.7 15.5
20.0 1.7
31.7 1.5

1,388.7 14.8

a-tocopherol
a-tocotrienol
g-tocopherol
g-tocotrienol
d-tocopherol
d-tocotrienol

Total (mg/kg)

1,532.7 24.0

48.3 4.5
152.7 5.8
90.7 2.1
1,235.0 6.1

CBOL

34.4 1.2

10.3 0.8
21.6 0.5

2.6 0.1

KWOL

2,112.5 108.9

407 22.9

1,640.7 86.9
7.2 0.3
58.3 3.2

RROL

280.3 17.5

260.3 13.7

20.0 3.8

STOL

BBOL = blackberry seed oil; BUOL = blueberry seed oil; CBOL = cranberry seed oil; KWOL = kiwi seed oil; RROL = red raspberry seed oil;
STOL = strawberry seed oil.

BUOL

BBOL

Tocopherol content (mg/kg)

TABLE 4.
TOCOLS (TOCOPHEROL AND TOCOTRIENOL) CONTENT AND COMPOSITION OF SEED OILS

BIOACTIVES OF BERRY SEEDS

45

46

V. VAN HOED ET AL.

Oxidative Stability
It is frequently reported that the oxidative stability of an oil is influenced
by its fatty acid composition, the oil quality parameters and tocopherol
content. This effect was most clearly observed for oils with highest and lowest
stability. KWOL, with the highest linolenic acid (C18:3) content, and the
lowest tocopherol content, exhibited the lowest oxidative stability (OSI), only
0.17 h at 97.8C (Table 1). Moreover, it had the highest PV, and the second
highest AnV (after STOL), thus resulting in the highest TOTOX value.
On the other hand, BBOL, having the lowest C18:3 content combined
with a high tocopherol content, showed the lowest PV and highest OSI (8.2 h).
The decreasing tocopherol content corresponded with a decrease in OSI
values, except for RROL, which was rich in tocopherols but is one of the least
stable oils. Although RROL did not contain the highest amount of C18:3, it
possessed the highest total PUFA content (C18:3 + C18:2), which may partly
explain its low OSI, and high PV, which was apparently not compensated by
the high content of tocopherols.
The OSI values of the studied berry seed oils were rather poor compared
with common vegetable oils with considerable PUFA content. Soybean oil and
corn oil, for example, showed OSI values of 16 and 28 h, respectively. These
findings are in agreement with the results of Parry et al. (2005), who reported
an oxidative stability of 2045 h at 80C for berry seed oils, compared with 47
and 66 h for soybean and corn oils, respectively. For comparison, the observed
OSI was estimated at 80C (Table 1). The results for berry seed oils were only
slightly lower, or comparable to those reported by Parry et al. (2005). There
was no literature data available for KWOL.
The low OSI values can be at least partially explained by the exceptionally high PUFA content. Thus, these oils require refrigerated storage in wellsealed small bottles, protected from light. Besides fatty acids, also tocopherols,
polyphenols, and some other antioxidants (pigments, squalene) or metal catalysts may contribute to the oxidative stability of the oil (data not reported).
CONCLUSIONS
There is a growing consumer awareness for healthy food products. In the
fat and oils industry this is reflected in a trend toward low trans and more
(poly)unsaturated fatty acids. Specialty oils, like berry seed oils, have a unique
fatty acid profile and they possess interesting minor components. They are also
produced from waste streams which makes them interesting from an economical point of view. The novel oils examined have nutritionally interesting fatty
acid profiles, tocol and phytosterol contents, but a rather low oxidative stability, which requires careful packaging and storage. Further studies are needed

BIOACTIVES OF BERRY SEEDS

47

to examine the oxidative stability and to characterize the oils polyphenol

content and radical scavenging capacities.
The characteristic chemical properties are likely to have an important role
to ensure the authenticity of these specialty, high-value oils.
ACKNOWLEDGMENTS
We sincerely thank Agnieszka Falkiewicz and Natalia Stelmaczonek,
Martien Segers from the lab of Food Technology and Engineering and Peggy
Dijckmans and Lieve Wozniak from Desmet-Ballestra for their efforts and
assistance during the course of this study.
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