Industrial Crops and Products 49 (2013) 130–135

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Industrial Crops and Products

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Compositional characteristics of sour cherry kernel and its oil as

influenced by different extraction and roasting conditions
Cemile Yılmaz, Vural Gökmen ∗
Department of Food Engineering, Hacettepe University, 06800 Beytepe, Ankara, Turkey

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

Article history: Sour cherry seeds arise as a waste material during processing of the fruits into processed products such
Received 11 December 2012 as canned or frozen sour cherry, and sour cherry juice. This study aimed to investigate the chemical
Received in revised form 19 April 2013 composition of the kernels in depth for potential utilization as a source of oil, protein and dietary fibers.
Accepted 27 April 2013
The kernel was found to contain 17.0% of oil, 29.3% of proteins and 30.3% of dietary fibers. Conven-
tional hexane and supercritical carbon dioxide (SC-CO2 ) were used to extract oil from the kernels. The
Keywords:
kernel oil was found to contain palmitic acid (6.4%), stearic acid (1.2%), oleic acid (46.3%), linoleic acid
Sour cherry seed kernel
(41.5%), and linolenic acid (4.6%). Extraction technique had no significant effect on fatty acid composi-
Oil extraction
Roasting
tion of kernel oil. The oil extracted by hexane contained significantly higher levels of tocopherols and
Bioactive compounds ␤-carotene than the oil extracted by SC-CO2 . The effect of ethanol used as a co-solvent in both extrac-
tion techniques on the composition of oil was determined. Using ethanol with both hexane and SC-CO2
increased total phenolic content, antioxidant capacity and ␤-carotene content of oil. Roasting kernels at
160 ◦ C for 30 min decreased total tocopherols (9.8%), but increased total phenolic content (4.5 times) and
hydroxymethylfurfural (1.4 mg/L) in resulting oil.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction by-product valorization. The effect of hexane and supercritical car-

Sour cherries, species of Prunus in the subgenus Cerasus,
are grown throughout the world at amounts about 1.2 million
tons/year (FAO, 2010). Sour cherries are mostly consumed as
processed products, such as canned and frozen sour cherry, or sour
cherry juice (USDA, 2012). During processing of sour cherries, high
amounts of seeds arise as a waste material that creates an important
disposal problem for the food industry.
Bak et al. (2006) have reported that sour cherry kernel extract
reduces ischemia reperfusion damage in isolated rat hearts. This
protective effect might be associated to bioactive compounds
in sour cherry kernels. The kernels contain anthocyanidins and
hydroxycinnamates (Bak et al., 2010). It was also reported that sour
cherry kernels comprise 32–36% oil, which is rich in ␥-sitosterol, ␤-
tocopherol and unsaturated fatty acids, with high content of oleic
acid (50–53%) and linoleic acid (35–38%) (Bak et al., 2010).
Due to their chemical properties, nutrients in sour cherry ker-
nel can pass into the oil at different percentages depending on
extraction methods and roasting process prior to oil extraction.
This study aimed to determine the compositional characteristics
of sour cherry seed kernel in depth for its potential utilization in
∗ Corresponding author. Tel.: +90 3122977108; fax: +90 3122992123.
E-mail address: vgokmen@hacettepe.edu.tr (V. Gökmen).
bon dioxide extractions, as well as use of ethanol as a co-solvent
during extraction was investigated on the chemical composition
of kernel oil. In addition, effect of various roasting conditions on
certain compositional characteristics of resulting oil was also deter-
mined.
2. Materials and methods
2.1. Chemicals and consumables
Potassium hydroxide, sodium carbonate, ␤-carotene, tetrahy-
drofuran, hydrochloric acid, boric acid, sodium hydroxide,
sulphuric acid, nitric acid, zinc sulphate, potassium hexacyano-
ferrate, glucose and amino acid standards were purchased from
Merck (Darmstadt, Germany). Ethanol, n-hexane, gallic acid,
Folin-Ciocalteu, potassium persulfate, ABTS reagent [2,2 -azino-
bis(3-ethylbenzothiazoline-6-sulphonic acid)] and fructose were
obtained from Sigma–Aldrich (Steinheim, Germany). Trolox [6-
hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid] and citric
acid monohydrate were purchased from Fluka Chemie AG (Buchs,
Switzerland). Disodium hydrogen phosphate, hydroxymethylfur-
fural (HMF) and sucrose were from Acros (Geel, Belgium). Formic
acid, acetonitrile, isopropanol and methanol were purchased from
J.T. Baker (Deventer, Holland). Amyloglucosidase and protease
were obtained from Novozymes (Bagsvaerd, Denmark). Mineral

0926-6690/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2013.04.048
 C. Yılmaz, V. Gökmen / Industrial Crops and Products 49 (2013) 130–135
standards were from Chem Lab (Zedelgem, Belgium). Ultra-pure
water was used throughout the experiments (Milli Q-System, Mil-
lipore, Milford, MA, USA).
Carrez I solution was prepared by dissolving 15 g of potassium
hexacyanoferrate in 100 ml of water and Carrez II solution was
prepared by dissolving 30 g of zinc sulfate in 100 ml of water.
Syringe filters (nylon, 0.45 ␮m), Acquity UPLC BEH C18 col-
umn (2.1 × 100 mm, 1.7 ␮m), Atlantis dC18 column (150 ×
2.1 mm, 3 ␮m), Atlantis dC18 column (250 × 4.6 mm, 5 ␮m) and
Atlantis HILIC column (150 × 2.1 mm, 3 ␮m) were purchased from
Waters (Millford, MA). HP-INNOWax column (60 m × 250 ␮m,
0.25 ␮m film thickness) was purchased from Agilent Technologies
(Waldbronn, Germany) and Shodex Sugar SH-1011 column (8.0
× 300 mm, 7 ␮m) was purchased from Showa Denko K.K (Tokyo,
Japan).
2.2. Sour cherry seeds
A local juice processing plant located in Mersin, Turkey provided
wasted sour cherry seed samples. These samples were evaluated
according to the flow sheet shown in Fig. 1. The kernels were sepa-
rated from the seeds by removing the outer shells prior to analyses
of proximate composition, sugars, vitamins, minerals, amino acids,
total phenolic content (TPC), and total antioxidant capacity.
2.3. Analysis of the proximate composition of kernels
Moisture (Method No: 925.10), ash (Method No: 930.05) and
protein (Method No: 984.13) contents were determined accord-
ing to Association of Official Analytical Chemists methods (AOAC,
1990). Total protein content was calculated by multiplying the
total nitrogen content by a factor of 6.25. Dietary fiber (Method
32-07.01) analysis was performed using American Association of
Cereal Chemists methods (AACC, 1991). To determine lipid con-
tent, ground kernels (200 mg) were extracted with hexane (3 ×
1 ml) and combined hexane phases were evaporated under a gen-
tle stream of nitrogen at 40 ◦ C. Carbohydrate content was calculated
by subtracting the total percentage of other components.
2.4. Analysis of sugars
Ground sample (1 g) was extracted with 1 ml of Carrez I, 8 ml
of hot water (70 ◦ C) and 1 ml of Carrez II solutions. After centrifu-
gation, upper aqueous extract was collected. Solid residue was
re-extracted with 8 ml and 7 ml of hot water, respectively and col-
lected extracts were centrifuged for 5 min at 11,180 × g. The 10 ␮l of
extract was injected into an Agilent 1100 High performance liquid
chromatography (HPLC) system (Waldbronn, Germany) consisting
of a quaternary pump, an autosampler, a refractive index detector
(RID) and a temperature controlled column oven. The chromato-
graphic separations were performed on a Shodex Sugar SH-1011
column using 0.01 mM H2 SO4 solution as mobile phase at a flow
rate of 0.7 ml/min at 25 ◦ C.
2.5. Analysis of water-soluble vitamins
Ground sample (1 g) was extracted three times with water (10,
5, 5 mL). After centrifugation at 7500xg for 5 min, supernatants
were collected. Water-soluble vitamins were determined by ultra-
performance liquid chromatography–tandem mass spectrometer
(UPLC-MS/MS) (Waters Corp., Milford, MA, USA). The chromato-
graphic separations were performed on an Acquity UPLC BEH C18
column using solvent A (0.1% formic acid in water) and solvent B
(0.1% formic acid in methanol) as the mobile phase at a flow rate
of 0.3 ml/min (40 ◦ C). The solvent gradient was programmed as fol-
lows: isocratic elution of 1% B and 99% A, 0–3 min; linear gradient
131
elution to 75% B and 25% A, 3–5 min; linear gradient elution to
1% B and 99% A, 5–5.1 min; and isocratic elution of 1% B and 99%
A, 5.1–8 min. The injection volume was set to 20 ␮l. The positive
ionization mode was selected for determination of thiamine, nico-
tinamide, pyridoxine, panthetonic acid and cyanocobalamin except
ascorbic acid. The source and desolvation gas temperature were
set at 120 ◦ C and 450 ◦ C; desolvation and cone gas (N2 ) flow rates
were 800 L/h and 10 L/h, respectively. The concentrations of vita-
mins were calculated by means of a calibration curve built in the
range between 1.0 and 10 mg/L.
2.6. Analysis of amino acids
Ground sample (100 mg) was weighed into a tube and
hydrolyzed with 4 ml of 6 N HCL at 110 ◦ C for 24 h. After hydroly-
sis, acid solution in the tube was diluted with water for 1000 times
and injected into an ultra high-performance liquid chromatogra-
phy (U-HPLC) Accela system (Thermo Fisher Scientific, San Jose,
CA, USA), directly interfaced to an Exactive Orbitrap MS (Thermo
Fisher Scientific, San Jose, CA, USA). Chromatographic separations
were performed on Atlantis HILIC column. Parameters of U-HPLC
and MS systems were performed as described by Gökmen et al.
(2012).
2.7. Analysis of minerals
Ground sample (1.0 g) was digested in a Mars Microwave Sys-
tem (CEM Corporation, Matthews) with 15 ml of nitric acid at 190 ◦ C
for 25 min applying the power of 1200 Watt. After digestion, sam-
ples diluted with water were filtrated from ashless filter paper and
injected into an atomic absorption spectrometer (Thermo Scientific
ICE 3000 Series, USA). The concentrations of calcium, sodium, mag-
nesium, potassium and iron minerals were calculated by means
of calibration curves built in the range between 0.0–25 mg/L,
0.0–1.0 mg/L, 0.0–0.2 mg/L, 0.0–1.0 mg/L and 0.0–2.0 mg/L, respec-
tively.
2.8. Analysis of TPC
Ground sample (250 mg) was hydrolyzed using 5 ml of 4 N NaOH
for 4 h at room temperature. After hydrolysis, the pH was adjusted
to 2.0 by 6 N HCl and hydrolysates were extracted with ethyl acetate
and diethyl ether (50:50, v/v) five times. The combined extracts
were evaporated by a gentle stream of nitrogen and the final residue
was re-dissolved in 1.5 ml of methanol-water (50:50, v/v). In addi-
tion, sour cherry kernel oils were extracted with methanol-water
(70:30, v/v) three times to measure TPC in oil samples.
TPC of oil samples and kernels were determined by using
Folin-Ciocalteu method. Extracts were mixed with Folin-Ciocalteu
reagent diluted with distilled water (1:10, v/v) and 20% aqueous
Na2 CO3 solution. After the mixture was allowed to stand in the
dark for 2 hrs, the tubes were centrifuged at 7155xg for 2 min
and the absorbance of supernatant was measured at 765 nm using
UV–vis spectrophotometer (Shimadzu, Kyoto, Japan). Standard cal-
ibration curve was prepared at different concentration of gallic acid
(0–100 ␮g/ml) to determine TPC as mg of gallic acid equivalent
(GAE) per liter of oil and 100 g of kernel.
2.9. Analysis of total antioxidant capacity
Antioxidant activity of the kernels was determined by
QUENCHER method described by Serpen et al. (2008) using ABTS.
Standard calibration curve was prepared at different concen-
trations of trolox (0–500 ␮g/ml) to determine total antioxidant
capacity as mmol Trolox equivalent antioxidant capacity (TEAC)
per 100 g of kernel. To determine total antioxidant capacity of sour
132 C. Yılmaz, V. Gökmen / Industrial Crops and Products 49 (2013) 130–135
Fig. 1. Scheme for potential utilization of sour cherry components.
cherry kernel oils, oil (100 ␮l) was weighed into a centrifuge tube
and the reaction was started by adding 10 ml of ABTS working solu-
tion. The mixture was shaken in the dark for 28 min and the tube
was centrifuged at 9200 × g for 2 min. After 30 min of reaction, the
absorbance of the supernatant was measured at 734 nm by UV–vis
spectrophotometer. The antioxidant capacity of samples was cal-
culated by using standard calibration curve as mmol TEAC per liter
of oil.
2.10. Extraction of oil from the seed kernels
The kernels were roasted in a temperature-controlled oven
(Memmert, Germany) at 160 ◦ C for 0 (control), 10, 20, 30, and
40 min in order to determine the effect of heating on the composi-
tion of resulting oil after extraction. The kernels were ground using
a coffee grinder prior to oil extraction by solvent or supercritical
carbon dioxide.
2.10.1. Solvent extraction
Solvent extraction was carried out using n-hexane with and
without ethanol (3.0%). 2 g of ground kernels was mixed with 5 ml
of solvent in a glass vessel. The mixture was shaken in an orbital
shaker for 15 min. The supernatant containing the oil was trans-
ferred into a glass flask. The residue was re-extracted twice by 5 ml
of solvent. Hexane in the combined extracts were evaporated at
40 ◦ C using a rotary vacuum evaporator. Oils were stored at −20 ◦ C
prior to analysis.
2.10.2. Supercritical carbon dioxide extraction
A Thar model 1000-2-FMC supercritical fluid extraction system
(Pittsburgh, PA, USA) equipped with an automated back pressure
regulator, a high pressure pump, a modifier pump and a heat
exchange module was used. Supercritical carbon dioxide with and
without ethanol (3.0%) was used for the extraction at 60 ◦ C and
300 bar. The ground kernels (100 g) placed in extraction vessel of
the system were extracted for 2 hrs at a total flow rate of 25 g/min.
Ethanol residues in resulting oils were evaporated by a gentle
stream of nitrogen. Oils were stored at −20 ◦ C prior to analysis.
2.11. Analysis of fatty acids in kernel oil
Oil (200 mg) was extracted with 10 ml of hexane and methyl-
ated with 100 ␮l of 2 N potassium hydroxide prepared in methanol.
After the resulting sample was centrifuged at 7500 × g for 2 min, the
upper phase was taken into a vial. 1 ␮l of sample was injected into
the gas chromatograph at a split ratio 1:40. Fatty acid composition
was determined by using an Agilent 7890A gas chromatogra-
phy (GC) system equipped with a capillary column (HP-INNOWax
19091N-136), an auto injector (Agilent 4513A) and a mass detec-
tor (Agilent VL MSD 5975C). The temperatures of injector port and
detector were operated at 250 ◦ C and 230 ◦ C. The column tempera-
ture was initially held at 60 ◦ C for 10 min and increased to 220 ◦ C at
rate of 4 ◦ C/min and held isothermal for 10 min and then increased
to 240 ◦ C at rate of 1 ◦ C/min. The flow rate of the carrier gas (helium)
was 1.5 ml/min. The analysis was performed by using electron-
impact ionization (70 eV) in scan mode. Famedb23 and NIST05a
libraries were used to identify fatty methyl esters and the results
were expressed as a percentage of fatty acids.
2.12. Analysis of tocopherols in kernel oil
Oil (1 ml) was extracted with 5 ml of the mixture of isopropanol-
methanol (50:50, v/v) three times. Combined extract was filtered
through 0.45 ␮m nylon filter prior to analysis. Tocopherols were
determined by using an Agilent 1200 HPLC system (Waldbronn,
Germany) consisting of a quaternary pump, an autosampler, a tem-
perature controlled column oven and equipped with an Agilent
6130 MS detector. Tocopherols were separated on Atlantis dC18
column (150 × 2.1 mm, 3 ␮m) using the mixture of methanol and
1.0% formic acid in water (95:5, v/v) at a flow rate of 0.3 ml/min
at 30 ◦ C. The injection volume was 10 ␮l. Data acquisition was per-
formed in SIM mode with positive ionization. Protonated molecular
ions ([M+H]+ ) having m/z 431.7 for ␣-tocopherol, 417.7 for ␤- and
␥-tocopherol and 403.7 for ␦-tocopherol were selected for iden-
tification and quantification of tocopherols. The concentrations of
tocopherols were calculated by means of calibration curves built in
the range between 1.0 and 100 mg/L.
 C. Yılmaz, V. Gökmen / Industrial Crops and Products 49 (2013) 130–135 133

2.13. Analysis of ˇ-carotene in kernel oil Table 1

Some compositional characteristics of sour cherry kernel (n = 3).

Oil (1 ml) was extracted with 5 ml of the mixture of isopropanol- Amount in 100 g seed kernel
methanol (50:50, v/v) three times. Combined extract was filtered Carbohydrates 46.62 ± 1.10 g
through 0.45 ␮m nylon filter prior to analysis. ␤-carotene was Sugars 2.91 ± 0.29 g
determined by using an Agilent 1200 HPLC system (Waldbronn, Sucrose 0.30 ± 0.00 g
Germany) equipped with a diode array detector (DAD). ␤-carotene Glucose 0.96 ± 0.05 g
Fructose 1.65 ± 0.25 g
was separated on Atlantis dC18 column (150 x 2.1 mm, 3 ␮m) using
Dietary fibers 30.25 ± 3.33 g
acetonitrile as the mobile phase at a flow rate of 0.3 ml/min at Lipids 17.00 ± 0.59 g
30 ◦ C. Chromatograms were recorded at 450 nm. The injection vol- Saturated fat 1.28 ± 0.05 g
ume was 10 ␮l. The concentration of ␤-carotene was calculated Monounsaturated fat 7.96 ± 0.03 g
Polyunsaturated fat 7.76 ± 0.05 g
by means of calibration curves built in the range between 1.0 and
Proteins 29.34 ± 2.20 g
10 mg/L. Amino acid composition (%)
Glutamic acid 27.96 ± 0.44
Arginine 9.30 ± 0.13
2.14. Analysis of HMF in kernel oil Proline 8.25 ± 0.18
Aspartic acid 7.55 ± 0.07
HMF content of kernel oil was determined according to the Phenylalanine 7.05 ± 0.13
Glycine 6.52 ± 0.00
method described by Durmaz and Gökmen (2010). An ultra fast Lysine 5.28 ± 0.02
liquid chromatography (UFLC) system (Kyoto, Japan) consisting Alanine 4.57 ± 0.32
of a quaternary pump, an autosampler, a diode array detector Serine 4.49 ± 0.20
(DAD), and a temperature-controlled column oven was used. The Ash 3.11 ± 0.49 g
Potassium 448.41 ± 2.05 mg
chromatographic separations were performed on an Atlantis dC18
Sodium 15.24 ± 2.16 mg
column (250 × 4.6 mm, 5 ␮m) using the isocratic mixture of 10 mM Calcium 275.13 ± 0.50 mg
aqueous formic acid solution and acetonitrile (90:10, v/v) at a flow Iron 4.48 ± 0.44 mg
rate of 1.0 ml/min at 25 ◦ C. Data acquisition was performed by Magnesium 234.39 ± 2.00 mg
recording chromatograms at 285 nm. The concentration of HMF Vitamin
Water soluble
was calculated by means of a calibration curve built in the range
Vitamin B1 0.15 ± 0.11 mg
between 0.1 and 1.0 mg/L. Vitamin B6 0.18 ± 0.03 mg
Vitamin B5 0.03 ± 0.00 mg
Vitamin B3 0.57 ± 0.02 mg
2.15. Statistical analysis Lipid soluble
Vitamin E 6.36 ± 0.81 mg
Total phenolic content 53.87 ± 0.02 mg GAE
The results were reported as mean ± standard deviations. Sig-
Total antioxidant capacity 0.92 ± 0.08 mmol TEAC
nificant differences (p < 0.05) were evaluated by Duncan’s multiple Moisture 3.91 ± 0.11 g
range test after the analysis of variance (ANOVA), by using SPSS
The results are given as mean ± S.D. GAE refers to gallic acid equivalent. TEAC refers
13.0 statistical package. to trolox equivalent antioxidant capacity.

3. Results and discussion
3.1. Compositional characteristics of sour cherry kernels
Table 1 gives compositional characteristics of sour cherry ker-
nels. The kernels were found to be rich in carbohydrates, proteins
and lipids with their corresponding concentrations of 46.6%, 29.3%,
and 17.0%, respectively. Dietary fibers were determined as the
abundant form of carbohydrates (30.25%) whereas sugars were
in relatively low quantities (2.91%) in the kernels. Therefore sour
cherry kernels have importance from the point of dietary fiber
content which is related to the intestinal regulation, the intestinal
absorption of glucose and reduction of cholesterol levels (Rodriguez
et al., 2006). The results revealed that glutamic acid was found
as the dominant amino acid with its percentage contribution of
27.96%. In addition, lysine, a limiting essential amino acid in most
cereals, was found in relatively high quantity in sour cherry kernel
with its percentage contribution of 5.28%. Potassium and calcium
were major minerals found in sour cherry kernel. However, mineral
content obtained in this study was more higher than that obtained
by Popa et al. (2011). According to results of water-soluble vitamin
analyses, sour cherry kernel contained vitamin B1 , B3 , B5 and B6
whereas vitamin B12 and ascorbic acid were not detected in ker-
nel. Relatively high oil content (17.0%) of sour cherry kernel makes
it a valuable source of edible oil when compared with oil content
of other oil seeds and tree fruits such as soybean (18–20%), corn
(3.1–5.7%), sunflower (35–45%), and olive (15–35%) (O’Brien, 2004).
It has been previously reported that sour cherry kernels comprise
32–36% of oil (Bak et al., 2010). The composition of the kernels from
different varieties or locations may differ significantly.
3.2. Chemical composition of sour cherry kernel oil
Table 2 gives the fatty acid compositions of kernel oils extracted
by organic solvent (hexane or ethanol modified hexane) and by
supercritical fluid (SC-CO2 or ethanol modified SC-CO2 ). It was
found that sour cherry kernel oil composed of mainly polyunsat-
urated fatty acids in the form of linoleic and linolenic acids, and
monounsaturated fatty acid in the form of oleic acid. The sum of
saturated fatty acids (stearic acid and palmitic acid) was less than
10% in the kernel oils. Popa et al. (2011) reported that sour cherry
kernel oil contained low levels of arachidic acid (0.9%) and myristic
acid (0.5%) while dominant saturated fatty acids were palmitic acid
(11%) and stearic acid (6.4%).
The sum of linoleic and linolenic acids was found to exceed
45% in the kernel oils. Linoleic and linolenic acids, classified as ␻-6
and ␻-3 polyunsaturated fatty acids respectively, form an impor-
tant part of human diet, because lack of them in the diet creates
adverse effects on human health such as cardiovascular disease
and skin lesion (Temelli et al., 2007). Except palmitic acid, there
was no significant difference (p > 0.05) between fatty acid compo-
sitions of the kernel oils extracted with SC-CO2 and hexane. It was
also found that using ethanol with both hexane and SC-CO2 did
not exhibit any change in the fatty acid compositions of the ker-
nel oils (p > 0.05). Similar results have been previously reported
in the literature. Vagi et al. (2007) reported that tomato pomace
134 C. Yılmaz, V. Gökmen / Industrial Crops and Products 49 (2013) 130–135

Table 2
Composition of sour cherry kernel oils obtained by different extraction methods.

Solvent extraction Supercritical extraction

Hexane With ethanol SC-CO2 With ethanol

Fatty acids (%)

C16:0 Palmitic acid 6.23 ± 0.15a 5.93 ± 0.16a 7.24 ± 0.16b 6.14 ± 0.12a
C18:0 Stearic acid 1.33 ± 0.13a 1.08 ± 0.17a 1.33 ± 0.12a 1.19 ± 0.01a
C18:1 Oleic acid 46.80 ± 0.16a 47.94 ± 1.33a 44.99 ± 1.38a 45.54 ± 1.33a
C18:2 Linoleic acid 40.58 ± 0.13a 41.23 ± 1.04a 41.81 ± 0.13a 42.26 ± 1.17a
C18:3 Linolenic acid 5.06 ± 0.14a 3.80 ± 1.107a 4.63 ± 1.098a 4.86 ± 1.25a
Total tocopherols (mg/L) 428.62 ± 2.67a 376.56 ± 3.59b 312.15 ± 3.04c 381.68 ± 2.23b
␣-tocopherol 65.66 ± 2.77a 61.48 ± 2.80a 89.47 ± 3.49b 94.93 ± 2.44b
(␤+␥)-tocopherol 274.06 ± 2.91a 237.06 ± 1.64b 183.80 ± 2.54c 228.25 ± 2.47d
␦-tocopherol 88.89 ± 2.81a 78.02 ± 2.43b 38.88 ± 2.8c 58.50 ± 2.26d
␤-carotene (mg/L) 8.47 ± 0.20a 10.03 ± 0.27b 5.65 ± 0.21c 6.00 ± 0.11c
Total phenolic content (mg GAE/L) 6.60 ± 0.41a 9.61 ± 0.2b 19.34 ± 0.00c 27.87 ± 0.90d
Antioxidant activity (mmol TEAC/L) 1.44 ± 0.18a 2.20 ± 0.13c 2.06 ± 0.13b 2.23 ± 0.15c

The results are given as mean ± S.D. Different letters within the same row indicates statistical significance (P < 0.05). GAE refers to gallic acid equivalent. TEAC refers to trolox
equivalent antioxidant capacity.

extracts obtained by n-hexane, ethanol and SC-CO2 solvents had
same fatty acid compositions. Sanchez-Vicente et al. (2009) also
did not observe any significant difference in the fatty acid compo-
sition among the peach seed oils obtained using SC-CO2 , ethanol
modified SC-CO2 and hexane extraction.
Besides responsibility for the color of many foods, carotenoids
play a crucial role in human diet as precursors of vitamin A and
as antioxidants (Pereira et al., 2010). As reported in Table 2, ␤-
carotene content ranged from 5.65 mg/L in the SC-CO2 extract to
6.00 mg/L, 8.47 mg/L and 10.03 mg/L in ethanol modified SC-CO2 ,
hexane and ethanol modified hexane extracts, respectively. The
kernel oil extracted with hexane had significantly higher amount
of ␤-carotene than that extracted with SC-CO2 (p < 0.05). Addi-
tion of ethanol to hexane increased significantly the recovery of
␤-carotene from kernel to oil (p < 0.05), whereas the difference
between pure SC-CO2 and ethanol modified SC-CO2 extracts in
terms of ␤-carotene content was not statically significant (p > 0.05).
Eller et al. (2010) reported that ␤-carotene concentration in tomato
seed oil was highest for the hexane accelerated solvent extraction
(ASE) and ethanol ASE treatments with a very low content for the
SC-CO2 extraction. It was reported by Temelli et al. (2007) that sol-
ubility in SC-CO2 depends on fluid density and properties of solute
such as molecular weight and polarity. Therefore, it is obvious that
␤-carotene can be extracted by hexane and SC-CO2 because of being
a non-polar compound.
Nuts and oilseeds are rich natural sources of tocopherols act-
ing as chain breaking antioxidants and protectors of unsaturated
fatty acids against oxidative damage (Güçlü-Üstündağ and Temelli,
2004). Four tocopherol isomers (␣-, ␤-, ␥- and ␦-tocopherols) were
determined in sour cherry kernel oils. As reported in Table 2, total
tocopherol concentration of the kernel oils extracted with different
solvents ranged from 312.15 mg/L to 428.62 mg/L. These levels are
comparable to olive oil (110 ± 40 mg/L), palm oil (240 ± 60 mg/L)
and peanut oil (482 ± 345 mg/L) (O’Brien, 2004).
Total tocopherols in the hexane extract were significantly higher
than in the SC-CO2 extract (p < 0.05). On the other hand, ␣-
tocopherol concentration of the hexane extract (65.66 mg/L) was
significantly lower than that of the SC-CO2 extract (89.47 mg/L)
(p < 0.05). Using ethanol caused significant differences in total toco-
pherols for both hexane and SC-CO2 extraction. When ethanol
was used, total tocopherols decreased for hexane extraction, but
increased for SC-CO2 extraction (p < 0.05).
TPC of the kernel oils ranged from 6.60 mg GAE/L to
27.87 mg GAE/L in the following order: hexane < hexane with 3%
ethanol < SC-CO2 < SC-CO2 with 3% ethanol extracts. Using ethanol
with both hexane and SC-CO2 significantly increased TPC content of
the kernel oil (p < 0.05) due to high polarity of ethanol facilitating
the extraction of phenolic compounds from the kernels. In gen-
eral, SC-CO2 was better than hexane with respect to extraction of
phenolic compounds.
Similar to TPC, the antioxidant activity was highest in oil
extracted with ethanol modified SC-CO2 (2.23 mmol TEAC/L). This
result was similar to the report that the SC-CO2 extracts of black
sesame seeds exhibited significantly higher antioxidant activities
comparable to that by n-hexane extraction (Hu et al., 2004).
3.3. Roasting effect on chemical composition of sour cherry kernel
oil
Roasting at 160 ◦ C for different times had no significant effect
on fatty acid composition of sour cherry kernel oil (p > 0.05). This
result is in accordance with a previous report that there were no
differences in fatty acid composition of rice germ oils prepared at
different roasting temperatures and times (Kim et al., 2002).
Fig. 2 shows the changes in some chemical characteristics of sour
cherry kernel oil with roasting time. The concentration of total toco-
pherols tended to decrease linearly as the roasting time increased.
The results of previous studies to determine the effect of roasting
on tocopherols are contradictory. Decrease in the concentrations
of tocopherols of oil samples was attributed to oxidation reactions
taking place during roasting process (Murkovic et al., 2004). It was
also reported by others that roasting caused an increase in toco-
pherols because of better extraction of destroying cell walls (Lee
et al., 2004).
It was observed that HMF concentration increased gradually as
the roasting time increased. The oil extracted from raw sour cherry
kernels has no HMF expectedly. However, roasting of the kernels
induces the Maillard reaction forming HMF as an intermediate com-
pound. It was previously reported that furfurals formed in oilseeds
during roasting could pass to oil during extraction due to their rel-
atively less polarity (Durmaz and Gökmen, 2010). Only trace levels
of HMF were determined in the oils extracted from the kernels
roasted at 160 ◦ C for 10 and 20 min. Longer roasting times caused a
significant increase in HMF concentrations of resulting kernel oils
(p < 0.05).
Roasting affected also TPC of sour cherry kernel oils. The TPC of
unroasted sour cherry kernel oil was 9.18 mg GAE/L. It increased
linearly as the roasting time increased. Moreover, total antioxidant
capacity of the kernel oil increased linearly reaching to a plateau
value within 20–30 min of roasting at 160 ◦ C. It began to decrease
afterward.
Roasting forms various neo-formed compounds having antiox-
idant activity. In addition, it may increase the extractability of
several naturally occurring antioxidants by modifying the food
 C. Yılmaz, V. Gökmen / Industrial Crops and Products 49 (2013) 130–135
Fig. 2. Change of some chemical characteristics of sour cherry kernel oil with roasting time. (a) Total phenolic content, (b) HMF, (c) Antioxidant activity, (d) Total tocopherols.
matrix. In the meantime, some heat-labile antioxidant compounds
are lost during extended heat treatment. From the viewpoint
of total antioxidant capacity of oil, there is a balance between
neo-formed compounds and degradation of naturally occurring
antioxidants in roasted kernels. This kind of behaviour was pre-
viously reported for nuts and oilseeds (Açar et al., 2009). In overall,
the results suggest that a mild heat treatment of sour cherry seed
kernel is useful to improve the bioactive profile of its oil without
increasing the concentration of undesired compounds like HMF.
4. Conclusion
The results revealed that sour cherry kernel might be utilized
as a source of dietary fibers, proteins and lipids. Comparing to
other edible oils from various plants, the oil extracted from sour
cherry seed kernel either by hexane or SC-CO2 is rich in certain
bioactive compounds like polyunsaturated fatty acids, tocopherols,
␤-carotene and phenolic compounds. Further improvements in the
compositions of bioactive compounds in the resulting kernel oil can
be achieved by fine tuning the extraction and roasting conditions.
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