This article appeared in a journal published by Elsevier.

The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/copyright
 Author's personal copy

Food Chemistry 124 (2011) 983–990

Contents lists available at ScienceDirect

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

Characteristics of Prunus serotina seed oil q

Itsi Alveano Aguerrebere a, Alejandra Rojas Molina a, B. Dave Oomah b,*, John C.G. Drover b
a
PROPAC, Programa de Posgrado en Alimentos del Centro de la Republica, Laboratory of Chemical and Pharmacological Natural Products Research, School of Chemistry,
Universidad Autónoma de Querétaro, Querétaro, Qro. 76010, Mexico
b
National Bioproducts and Bioprocesses Program, Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, Summerland, British Columbia, Canada V0H 1Z0

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

Article history: Oils from Prunus serotina raw and toasted seeds extracted with hexane and supercritical CO2 were eval-
Received 29 April 2010 uated for their physicochemical characteristics. Supercritical CO2 extracted the least oil (21.3%), with high
Received in revised form 21 May 2010 absorbing carotenoid pigments. P. serotina oil had characteristically high refractive index and density
Accepted 14 July 2010
with three typical absorbance peaks in the UVC (100–290 nm) range centred at 260, 270 and 280 nm.
The oil was highly polyunsaturated and abundant in oleic (35%), a-elostearic (27%), linoleic (27%), pal-
mitic (4%), stearic (4%) and b-elostearic (1%) acids. P. serotina seed oil exhibited at least three distinct
Keywords:
thermal structural transitions between 35 and 13 °C with two reversing transitions between 19
Prunus
Prunus serotina seed oil
and 12 °C. Thermal oxidation by differential scanning calorimetry (DSC) revealed a three step oxidation
Oil quality of P. serotina seed oil with the mean onset and oxidation temperatures at 121 and 130–273 °C, respec-
FT-IR tively, depending on processing. Supercritical CO2 extraction and toasting significantly affected the ther-
DSC mal and oxidation characteristics, fluorescence, and fatty acids of oils.
Fluorescence Crown Copyright Ó 2010 Published by Elsevier Ltd. All rights reserved.
Black cherry
Capulin
Chemical and physical parameters
Almond
Supercritical CO2

1. Introduction P. serotina fruits are used in Mexican traditional medicine for

Prunus serotina (Rosaceae) is a 60–90 foot-tall native North
American tree, widely distributed in Mexico, commonly called
‘‘capulín”, or American black cherry (Ibarra-Alvarado et al., 2009).
The bark has been used by the Iroquois, Ojibwa, Malecite, and Del-
aware indigenous people of the North American boreal forest re-
gions of Canada for diabetes related symptoms (McCune & Johns,
2007), and traditionally for the treatment of inflammatory-related
symptoms, such as cold, fever, and sore throat. It is known to exert
anticancer properties and antiproliferative effects via down-
regulation of cyclin D1 expression in human colorectal cancer cells
(Mazzio & Soliman, 2009; Yamaguchi, Liggett, Kim, & Baek, 2006).
In Guatemala the decoction of Prunus capuli Cav. (Rosaceae) leaves
is used for gastrointestinal disorders, such as diarrhoea, dysentery,
colic and inflammation, probably due to the strong antibacterial
activity demonstrated in leaf extract (Cáceres et al., 1993). P. sero-
tina is considered as one of the plants containing compounds that
exhibit anxiety reducing activity, allowing its use as a sleep induc-
ing agent (Yurchak, Dobberstein, & Heist, 2003).
q from the leaves of P. serotina, promoted vascular smooth muscle
Pacific Agri-Food Research Centre.
* Corresponding author. Tel.: +1 250 494 6399; fax: +1 250 494 0755.
E-mail address: oomahd@agr.gc.ca (B.D. Oomah).
the treatment of diarrhoea and cough. These fruits are also part
of the Mexican diet, and are consumed fresh, dried or prepared
in jam (Ibarra-Alvarado et al., 2009). Components of the P. serotina
seed have not received much attention and are still poorly docu-
mented, although those of other prunus species have been investi-
gated. For example, black cherry kernel, a waste product from fruit
juice processing contains 20% oil, consisting mainly of linoleic
(77%), linolenic (11%), and palmitic (8%) fatty acids. The oil is used
in the preparation of salads and cosmetics in the United States and
of drugs in traditional medicine in Iran. The oil is also highly effec-
tive in the production of the antibiotic erythromycin (Hamedi,
Malekzadeh, & Niknam, 2002; Hamedi, Malekzadeh, & Saghafi-
nia, 2004). Another cherry (Prunus cerasus) seed oil (30–33% ker-
nel) with no detectable cyanogens exhibiting strong antioxidant
activity, has been proposed for cosmetic and drug use (Tósaki
et al., 2008).
As a part of a study directed towards the screening of plants
used in Mexican traditional medicine for the treatment of cardio-
vascular diseases, our research group found that the lyophilised
aqueous and methanolic extracts, and the essential oil obtained
relaxation of the rat aorta (Ibarra-Alvarado et al., 2009). The health
benefits of P. serotina leaves for the treatment of hypertension was

0308-8146/$ - see front matter Crown Copyright Ó 2010 Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.07.040
 Author's personal copy

984 I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990

attributed partly to the presence of oil components (Ibarra- were measured with a spectrophotometer (SPECTRAmax Plus
Alvarado et al., 2009). The present study aimed at evaluating the 384, Molecular Devices Corp., Sunnyvale, CA). The colour of the oils
characteristics of P. serotina seed oil that may have beneficial was also compared with colour charts and the codes of the Mun-
health effects and changes incurred by processing. This is a contin- sellÒ Book of Color (New Windsor, NY). The refractive index was
uation of our studies on the utilisation of horticultural crops for measured with an Abbe Mark II refractometer.
innovative uses, leading to the development of new products for The fluorescence of oil was measured with a microplate spec-
the functional food, cosmetic, therapeutic and nutraceutical trofluorimeter (SPECTRAmax GEMINI EM, Molecular Devices Corp.,
industries. Sunnyvale, CA). Excitation at 365 nm gave excellent results and
was used throughout the study. The emission fluorescence spectra
were recorded from 420 to 850 nm. A cut off filter at 420 nm was
2. Materials and methods
used to increase the sensitivity and block the light below the cut
off wavelength, using 200 ll aliquot for analysis.
P. serotina seeds were obtained from fruits picked on May 2009
The FT-IR spectra of the oils were recorded at 22 °C using a
from Huejotzingo, Puebla, Mexico, washed and frozen to 70 °C
Nicolet 380 spectrometer (Thermo Electron Corp., Madison, WI)
(Revco PLUS Ultra Low Temperature Upright Freezers, Thermo Sci-
with SMART iTR diamond attenuated total reflectance (ATR acces-
entific). The seeds were removed from the pulp with plastic knives,
sory), with a 45° aperture angle generating a single bounce. The FT-
washed and allowed to dry. The seeds were cracked open with a
IR was equipped with a deuterated triglycine sulphate (DTGS)
mortar and pestle and the ‘‘almonds” transported to Summerland
detector, scanning over the frequency range of 4000–400 cm 1,
(BC, Canada), and stored in a desiccator prior to sample prepara-
at a resolution of 4/cm. Spectra were collected by using a rapid
tion. Toasting was performed in a comal (traditional Mexican
scan software running under EZOMNIC 8.0.342 (Nicolet, Madison,
cast-iron griddle pan), until the colour changed to slightly pink,
WI), and the spectrum for each sample was calculated from the
the seeds were cracked open with pliers, and then the ‘‘almonds”
average of 32 repetitive scans. A single drop of oil was placed di-
were stored in a desiccator prior to analysis. Commercial almond
rectly onto the ATR crystal and compressed using a standard pres-
oil (Spectrum Naturals, Hain Celestial Canada, Delta, BC) and
sure tower equipped with a pointed flat tip before the spectra
‘‘Whole natural almonds” (Trophy Foods Inc., Calgary, AB), pur-
collection. The spectra were collected in triplicate for each extract.
chased from a local food store, were used as controls. The almond
The thermal characteristics of P. serotina seed oil were mea-
was ground and extracted with hexane, as described below for P.
sured using a modulated differential scanning calorimeter (Modu-
serotina seeds.
lated DSC-2910, TA Instruments, New Castle, DE). A flow of
Samples (25 ± 0.5 g) were cryomilled in a 6800 SPEX Freezer/
nitrogen gas (145 ml/min) was used in the cell cooled by helium
Mill (SPEX, Metuchen, NJ 08840, USA) with a 5 min precooling per-
(145 ml/min) in a refrigerated cooling system. The instrument
iod and a two cycle program, consisting of a 1.5 min grinding time,
was calibrated for temperature and heat flow with mercury (melt-
followed by 1 min cooling between grindings, and an impactor
ing point, mp = 38.83 °C, TA Instruments standard), and succino-
speed of 14 s 1.
nitrile (mp = 58.06 °C, Fluka Chemie AG) and indium (mp =
The oil from all milled samples was extracted using hexane, as
156.6 °C, DH = 28.71 J/g, Aldrich Chemical Corp.). Oil samples (4–
described by Oomah, Mazza, and Przybylski (1996). Briefly, the
5 mg) were weighed in open solid fat index (SFI) aluminium pans
sample (25 g) was stirred for 2 h at 4 °C with hexane (75 ml). The
(T70529, TA Instruments). An empty pan was used as reference.
solvent was removed by vacuum filtration and the sample was fur-
The sample and reference pans were placed inside the calorimeter
ther extracted twice (2 h + 1 h). After the last filtration, the extracts
and kept at 70 °C for 5 min. The temperature was lowered from 70
were pooled, the hexane removed (vacuum rotary evaporation,
to 65 °C at a rate of 1 °C/min with modulation at a period of 60 s,
30 °C), purged with nitrogen, and stored at 30 °C until analysis.
and a temperature amplitude of 0.16 °C. Samples were then kept at
Extractions were performed in triplicate, duplicate for the toasted
65 °C for 1 min, and then raised again at the same rate, up to
samples, and analysed separately.
70 °C. Scans were performed at 1 °C/min. For thermal oxidation,
Supercritical CO2 extraction was carried out using 50 g milled
the pans were cooled to 10 °C and scanned by heating at 10 °C/
samples, as described by Athukorala, Mazza, and Oomah (2009),
min to 350 °C in the presence of oxygen (100 ml/min). The thermal
without co-solvent, for 3 h in a laboratory-scale supercritical fluid
oxidation measurements were performed in triplicate.
extraction system (Thar Technologies Inc., Pittsburgh, PA), with
The lipids were esterified in an alkaline medium (methanolic
carbon dioxide (99.95% purity, Praxair, Edmonton, AB) compressed
1 M KOH) as described previously (Beveridge, Girard, Kopp, & Dro-
to 30 MPa with the vessel (500 ml) temperature controlled at
ver, 2005). The top layer was transferred into a small vial and ana-
70 °C. The milled sample was placed in the extraction vessel,
lysed for its fatty acid methyl esters on an Agilent 6890 gas
heated to the set temperature of 40 °C, which was controlled by
chromatograph (Agilent Technologies Inc., Wilmington, DE),
a thermostat, and was monitored using the computer. The lipid-la-
equipped with a model 7683 autoinjector and a split/splitless
den gas from the extractor was passed through a heated metering
injector with a split ratio of 10:1, a flame-ionisation detector,
valve where the supercritical CO2 was depressurised, and the ex-
and a 100 M SP-2560 fused-silica capillary column (0.25 mm i.d.,
tracted oil was collected in a stainless vessel, whilst CO2 was
Supelco, Oakville, ON). The column temperature was programmed
vented out. The extract was flushed with nitrogen for 24 h (to con-
from 140 to 240 °C at 4 °C/min, and the injector and detector tem-
stant weight), weighed and stored in the dark at 30 °C until
peratures were set at 260 °C. Helium was the carrier gas. The
analysis.
instrument was controlled with an Agilent ChemStation (version
B.04.01). The peaks were identified using internal (heptadecanoic
2.1. Analytical procedures acid) and external standards (Supelco 37 mix components,
10 mg/ml; tung oil, Sigma–Aldrich Canada Ltd., Oakville, ON). The
The spectroscopic indices, K232 and K270, in the UV region, were analysis was done in duplicate.
determined as outlined in the Standard Methods for the Analysis of The oils were subjected to thin-layer chromatography (TLC) on
Oils, Fats and Derivatives (International Union of Pure and Applied precoated 0.20 mm silica gel 60 plates with a fluorescent indicator
Chemistry, IUPAC, 1985). The absorbencies at 670, 610, 560, and UV254 (Macherey–Nagel Inc., Bethlehem, PA) using the solvent sys-
535 nm, relating to chlorophylls, and at 475, 448, and 414 nm, tem dichloromethane/methanol/acetic acid (40/10/1, v/v/v), as de-
relating to carotenoids, of a 10% (w/v) solution of oil in hexane scribed by Horseley and Meinwald (1981). Samples and standards
 Author's personal copy

I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990 985

were mixed with 80% ethanol. The supernatant (5 ll) was spotted (1.70–1.77) corresponding to the highest absorptivity (K270, Table
on the TLC plate which was subsequently developed and dried 1). This UV spectra (maxima at around 260, 270 and 280 nm),
with a heat gun. Visualisation was carried out under short indicating cis, trans, trans configuration has previously been de-
(254 nm) UV light followed by charring with 5% sulphuric acid in scribed in a-eleostearic acid containing oils (Dyer et al., 2002).
methanol to locate cyanogenic glycosides. Amygdalin and prunasin The difference in the oil absorbance between the untreated and
standards were obtained from Sigma–Aldrich, Inc. (St. Louis, MO, the toasted seeds was not significant, but was reflected in their cor-
USA). responding Munsell colours (5Y 8.5/10 and 3.75Y 8.5/12). The high
At least three determinations were made for all assays. Analysis absorption at approximately 428, 453 and 475 nm, reflecting carot-
of variance by the general linear models (GLM) procedure and enoid (412–482 nm) pigments (typical of beta-carotene absorption
means comparisons by Duncan’s test were performed according at 425, 450 and 477 nm), characterised the supercritical CO2 ex-
to Statistical Analysis System (SAS Institute, 1990). tracted oil, and differentiated it from the untreated hexane ex-
tracted oil. The spectrum of P. serotina oil showed significant
absorbance (1.2) in the 280–315 nm (UVB) range at high concen-
3. Results and discussion tration (10 g/l). High absorptivity at 270 nm may be due to high
diene conjugation. The differences at all other absorbances were
The oil yield from supercritical CO2 extraction (21.3%, Table 1) not significant amongst the samples. The P. serotina seed oil colour
was significantly (P < 0.05) lower than that extracted with hexane. and absorbance values were similar to that of the commercial al-
Similar lower recoveries (10–34%) with supercritical CO2 extrac- mond oil.
tion were previously reported for flaxseed oil (Barthet & Daun, A representative fluorescence spectrum of P. serotina seed oils
2002; Bozan & Temelli, 2002). Toasting, a process known to pro- (Fig. 2) reveals the presence of three major peaks; a broad peak
duce the unique flavour of many Mexican foods, improved the oil (430–465) with the highest intensity at 440 nm, except for toasted
yield marginally by 5%, without being statistical significant. Under seed oil, a medium peak at 676 nm, and a distinct shoulder peak at
the same conditions, hexane extracted significantly (P < 0.0005) 729 nm (Table 2). The peaks at 440 and 676 nm also present in veg-
higher (46%) oil from almonds, over twice the amount obtained etable oils may be ascribed to the presence of chlorophyll in the oil,
from P. serotina with supercritical CO2 extraction. The first, second according to Kyriakidis and Skarkalis (2000). The intensity of the
and third hexane extractions produced on average 87.2%, 10.3%, peaks at 440 nm corresponds to the values of conjugated trienes,
and 2.5% of the total oil, respectively, independent of the source K270 of 1.77 and 1.70 for the hexane extracted oils of the raw and
material. The density (0.910–0.914) of P. serotina oil was similar toasted seeds, respectively. This indicates that fatty acid oxidation
to that of mustard seed, olive, rapeseed, and wild apricot kernel products may also be responsible for the fluorescence at 440 nm, as
oils (Kaya, Kola, Ozer, & Altan, 2008; Padley, Gunstone, & Harwood, proposed by Kyriakidis and Skarkalis (2000). The low intensity
1994), and higher than the commercial (0.902) and hexane ex- peak with an emission at 729 nm may be attributed to chlorophyll
tracted (0.900) almond oils. The refractive index of P. serotina oil d, the red absorbing species, typical of the far red excitation of pho-
(1.4704) was similar to hexane extracted almond oils, and ex- tosystem II reduction (Jennings & Forti, 1975; Mimuro, Hirayama,
ceeded those of most common vegetable oils (1.400–1.478) (Pad- Uezono, Miyashita, & Miyachi, 2000). Processing induced the pro-
ley et al., 1994), fruit oils (1.464–1.471) (Yu, Parry, & Zhou, duction of fluorescence pigments, as measured from the increase
2006), or wild apricot kernel oil (Kaya et al., 2008), probably in oil fluorescence at 676 and 729 nm during toasting, and at 440
reflecting a high degree of saturation, or the presence of substantial and 460 nm for supercritical CO2 extraction. The significant in-
amounts of conjugated double bonds. The conjugated diene value crease at 676 nm, the yellow spectrum characteristics of chloro-
of the oils (4.65–5.05) was considerably high, but not significantly phyll a, probably originates from the unbound or uncoupled
different amongst the samples. The triene values (4.91–5.42) of the chlorophyll (Kovacevic, Dewez, & Popovic, 2007). The hexane ex-
oils also exceeded those evaluated previously in our laboratory. tracted P. serotina seed oils had significantly higher fluorescence
The P. serotina seed oil has three characteristic absorbance peaks intensity at 440, 676, and 729 nm than the hexane extracted al-
in the UVC (100–290 nm) range centred at 260, 270 and 280 nm mond oil. The exceedingly high fluorescence observed for the com-
(Fig. 1). The central peak at 270 nm is of maximum intensity mercial almond oil at 440 and 460 nm may be associated with lipid
modification due to oxidation (Gatellier, Santé-Lhoutellier, Por-
tanguen, & Kondjoyan, 2009).
Table 1 The FT-IR analysis revealed that P. serotina seed oils had similar
Quality characteristics (colour) of P. serotina seed oils. absorbencies indicating similar structural features (Fig. 3), unaf-
Quality Hexane extracts SFE fected by processing. The spectra showed typical absorption bands
characteristics
Untreated Toasted Untreated in the 2923–2853, 1740–1745, 1120–1160, 1000–900, and 700–
725 cm 1 regions. The absorbencies at these spectral regions are
Oil yield 30.9a 32.5a 21.3b
Density 0.914a 0.913a 0.910a
generally associated with aliphatic groups (2923–2853), aldehydes
R index 1.491ab 1.491a 1.490b and ketones (1740–1745), etheric bands (1120–1160), cis to trans
Diene 5.050a 4.790a 4.650a isomerisation (1000–900), and disappearance of the cis double
Triene 5.423a 4.905a 5.050a bonds (700–725) (Beltrán Sanahuja, Prats Moya, Maestre Pérez,
K232 0.256a 0.270a 0.274a
Grané Teruel, & Martín Carratalá, 2009; Durmaz, Karabulut, Topçu,
K270 1.765a 1.701a 1.698a
A412 0.035a 0.036a 0.040a Asiltürk, & Kutlu, 2010; Russin, van de Voort, & Sedman, 2004). The
A428 0.0350b 0.0355ab 0.0395a P. serotina seed oils had a characteristic doublet stronger at 990
A453 0.033b 0.0335ab 0.0375a and weaker at 960 cm 1, probably suggesting the presence of con-
A475 0.034b 0.034b 0.039a
jugated trans fatty acids (Russin et al., 2004). The specific absorp-
A535 0.032a 0.033a 0.037a
A560 0.031a 0.032a 0.036a
tion band 960/990 cm 1 parallels the spectral feature 958/
A610 0.031a 0.032a 0.035a 987 cm 1, which is strongly expressed in oils with conjugated
A670 0.032a 0.032a 0.036a cis–trans–trans bonds (Spitzer, 1999) and consistent with the pres-
a–b
Means in a row followed by the same letter are not.
ence of a-eleostearic acid (9 cis, 11 trans, 13 trans-conjugated lin-
Significantly different by Duncan’s multiple range test at the 5% level. SFE, super- olenic acid) observed in the commercial tung oil. The spectra
critical fluid extract. observed between 1460 and 760 cm 1 and 3000 and 2800 cm 1
 Author's personal copy

986 I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990

2.0

Hex SFE Toasted

UV Absorbance 1.6

1.2

0.8

0.4

0.0
210 230 250 270 290 310 330
Wavelength (nm)

Fig. 1. Ultra violet visible spectra of oils at 0.03 g/l of hexane from P. serotina seeds.

1000
Hex
SFE
Toasted
800 Alm Hex

600
RFU

400

200

0
420 470 520 570 620 670 720 770
Wavelength (nm)
Fig. 2. Comparison of scanning fluorescence spectra of oils from P. serotina seeds and almond, at 365 nm excitation.

have previously been assigned by Tósaki et al. (2008) to the long

Table 2 carbonyl chain components of sour cherry seed oil.
Fluorescence of P. serotina seed oils at various emission wavelengths.a The P. serotina seed oil exhibited at least three distinct thermal
Sample Emission wavelength (nm) structural transitions between 43 and 6 °C. Two reversing tran-
440 460 676 729 sitions (between 19 and 12 °C), indicative of crystalline melting,
were observed corresponding to the a and b polymorphic forms,
P. serotina hexane 459c 449c 281b 120b
P. serotina hexane toasted 438c 398c 947a 181a
respectively (Fig. 4). The reversing component of the heat flow
P. serotina SFE 807b 830b 93c 84bc was highly sensitive to the toasting treatment (Table 3). For exam-
Almond hexane 297d 383c 55c 64c ple, the first endothermic peak (the a-form) of the oil from toasted
Almond commercial 5576a 5700a 45c 71c seed occurred at significantly higher temperature and heat flow
a
Means in a column followed by the same letter are not significantly different by than that of the untreated and supercritical CO2 extracted oils. This
Duncan’s multiple range test at the 5% level. SFE, supercritical fluid extract. surge in heat flow of the a-form resulted in a significant decrease
 Author's personal copy

I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990 987

0.30

Hex SFE Toasted Alm Com Alm Hex

0.25

0.20
Scale

0.15

0.10

0.05

0.00
3400 3000 2600 2200 1800 1400 1000 600
Wavenumber cm-1
Fig. 3. FT-IR spectral analysis of P. serotina seed oils.

-0.04
Heat Flow (W/g)

-0.08

-0.12
Hex

SFE

-0.16 Toasted

Alm Hex

Alm Com
-0.2
-35 -25 -15 -5 5
Temperature (°C)

Fig. 4. Modulated differential scanning colorimetry (MDSC) reversing component of P. serotina seed oils.

Table 3
Thermal characteristics of P. serotina seed oil.

Sample Reversing heat flow Nonreversing heat flow

T/ DH / Tb DH b T/ DH/ Tb DHb
P. serotina hexane 18.1b 26.5a 12.1ab 36.8b 42.0b 49.9b 5.5a 5.3a
P. serotina hexane toasted 17.4a 37.3b 11.5a 28.1a 39.7a 53.4a 5.4a 5.0a
P. serotina SFE 18.6b 25.3a 12.3b 37.8b 42.7c 48.8b 5.5a 4.9a
Almond hexane 16.2x 21.2x 9.7x 60.1x 36.7x 34.1x 9.4x 4.2x
Almond commercial 19.1y 19.9x 11.3y 61.6x 38.7y 48.6x 10.4y 3.4x

Means in a column (a–c and x–y, for P. serotina and almond, respectively), followed by the same letter are not significantly different by Duncan’s multiple range tests at the 5%
level. SFE, supercritical fluid extract.

in the second endothermic peak (the b-form), due to toasting. In as crystallisation, crystal perfection and reorganisation are as-
the nonreversing component curves to which kinetic events, such cribed, two exothermic and an endothermic peak were observed
 Author's personal copy

988 I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990

(between 43 and 40 °C), and around the 5.5 °C region. The
exothermic peak ( 43 to 40 °C with high activation energy of
49–53 J/g) suggests kinetic stability with high entropy, implying
a first-order transition. The second endothermic peak in the
5.5 °C region was assigned to the b crystallisation form. The val-
ues of the nonreversing enthalpies, DHa and DHb were not signif-
icantly different amongst the P. serotina seed oils, but different
from those of the almond oils (Table 3). The DHa for the nonrevers-
ing heat flow were approximately 1.4–1.9 times those of the
reversing values. The DHb values for the nonreversing and revers-
ing heat flow for P. serotina oils were significantly lower and
higher, respectively, than those of the almond oils. The DHa values
for the reversing heat flow of the almond oils were significantly
lower than those of the P. serotina seed oils, although the first tran-
sitions occurred at temperatures significantly different from those
of P. serotina.
The P. serotina seed oil exhibited two maxima on the DSC oxida-
tion curves, indicating that thermoxidation can be characterised by
at least two step exothermic effects (Fig. 5). These peaks could be
considered as an indication of the level of cross-linking. Oxidation
of P. serotina seed oil started at 117–124 °C, slightly below the tem-
peratures reported for edible oils (130–180 °C), and peaked at 135–
156 °C, depending on sample types (Fig. 5; Table 4). The supercrit-
ical CO2 extracted oil had the lowest onset, oxidation, peak 1 and
peak 4 temperatures, with the characteristic absence of peak 3. Sig-
nificant differences were observed in the onset and the peak 1 tem-
peratures amongst the P. serotina samples (P < 0.0001), with the oil
from the toasted seed showing the highest oxidation temperature.
The second peak at 250 °C was present only in the oil extracted
from toasted and by supercritical CO2, indicating instability at this
particular temperature. The fourth peak at 310–318 °C indicates
the inability of oxygen uptake, resulting in the complete thermal
polymerisation. The hexane extracted seed oil showed no signifi-
cant differences in the temperature of the fourth peak, an observa-
tion similar to that reported previously for hempseed and
Echinacea seed oils (Oomah, Busson, Godfrey, & Drover, 2002;
Oomah, Dumon, Cardador-Martinez, & Godfrey, 2006). The almond
oil showed higher thermo-oxidation temperatures than P. serotina
seed oils with significant (P < 0.0001) differences between hexane
and commercial oils.
The most abundant fatty acid in almond oil was oleic acid
(68.5%) followed by linoleic acid (22.9%), with smaller amounts of
palmitic (6%), stearic (1.5%) and palmitoleic (0.4%) acids (Table 5),
similar to those reported previously (Beltrán Sanahuja et al.,
2009). The fatty acid, except oleic, content of the hexane extracted
almond oil was significantly lower than that of the commercial oil.
The main fatty acids of the P. serotina seed oil was oleic (35.3%),
a-eleostearic (26.7%), linoleic (26.6%), palmitic (4.3%), stearic (4%),
b-eleostearic (1%), palmitoleic (0.1%) and a-linolenic acids (0.2%).
This fatty acid composition was within the range reported for the
oils of other Prunus species (Deineka, Gabruk, Deineka, & Manokh-
ina, 2002), but considerably different from those of the black cherry
kernel oil (77% linoleic, 11% linolenic, and 8% palmitic acids)
(Hamedi et al., 2002). Linoleic acid was less abundant, whereas
the a-eleostearic acid content was over twice the amount com-
pared to the cherry seed oils (Comes, Farines, Aumelas, & Soulier,
1992). The relatively low linoleic acid content observed in P. sero-
tina compared to the other cherry seed oils (37–47%), and particu-
larly the black cherry seed oil (77%), may probably be attributed to
the modification of linoleic acid at the D12 position to a-eleostearic
acid, and their inverse relationship throughout the seed develop-
ment (Dyer et al., 2002 and references therein). The doublet in

13
Hex SFE Toasted Alm Hex Alm Com
11

9
Heat Flow (W/g)

-1
100 150 200 250 300 350
Temperature (°C)
Fig. 5. Differential scanning colorimetry (DSC) of the thermo-oxidation profiles of P. serotina seed oils.

Table 4
Thermoxidation temperatures (°C) of P. serotina seed oils.a

Sample Onset Oxidation temperature Peak 1 Peak 2 Peak 3 Peak 4

P. serotina hexane 119.9b 128.4b 144.1c nd 267.9b 317.8a
P. serotina hexane toasted 124.4a 135.9a 155.7b 247.3b 277.0a 317.0a
P. serotina SFE 117.5c 124.7c 134.7d 249.5b nd 309.8b
Almond hexane 178.4x 184.4x 202.7a 272.9a 303.5x 331.3x
Almond commercial 174.4x 177.9y nd nd 295.9y 326.9x
a
Means in a column (a–c and x–y, for P. serotina and almond, respectively), followed by the same letter are not significantly different by Duncan’s multiple range test at the
5% level. nd, Not detected. SFE, supercritical fluid extract.
 Author's personal copy

I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990 989

Table 5
Fatty acid composition of P. serotina seed oil.

Sample Composition (mass %)

16:0 16:1 18:0 18:1 18:2 18:3 20:0 22:0 Unk a-ESA b-ESA
Hexane 4.26b 0.11 4.09b 35.15b 26.72b 0.18 0.40b 0.11 1.13b 26.96a 0.87
Toasted 4.15c 0.11 4.03c 35.32ab 26.15c 0.18 0.39c 0.11 1.20a 27.28a 1.09
SFE 4.44a 0.12 4.18a 35.61a 26.94a 0.20 0.41a 0.12 1.14b 25.77b 1.09
Almond commercial 6.10x 0.45x 1.54x 66.64y 24.29x 0.14x 0.08
Almond hexane 6.04y 0.42y 1.51y 69.18x 22.47y 0.12y 0.07

Means in a column (a–c and x–y, for P. serotina and almond, respectively), followed by the same letter are not significantly different by Duncan’s multiple range tests at the 5%
level. ESA, eleostearic acid. SFE, supercritical fluid extract.

the FT-IR spectrum with a strong band at 991 cm 1 and a weaker
band at 963 cm 1, and the UV spectral bands around 260, 269
and 280 nm, with the highest absorbance at 269 nm (Prashantha,
Premachandra, & Amarasinghe, 2009 and references therein), pro-
vides further confirmation for the presence of a-eleosteric acid.
Furthermore, a-eleosteric acid is specific to cherry, and detectable
only when esterification is carried out in an alkaline medium
(Comes et al., 1992) as in this study.
Processing significantly (P < 0.005) affected the fatty acid com-
position of the P. serotina seed oil (Table 5). The concentrations
of the major fatty acids, palmitic, stearic, oleic, and linoleic were
significantly higher in the oil extracted by supercritical CO2 than
hexane. This is contrary to the observation in the cherry seed oils,
where the fatty acid composition consisting of linoleic (40.8%),
oleic (32.6%), a-eleosteric (11.1%), and palmitic acids (5.3–6.0%),
was unaffected when extracted with hexane or supercritical CO2
(Bernardo-Gil, Oneto, Antunes, Rodrigues, & Empis, 2001). The oil
extracted from the toasted seed contained significantly lower con-
tent of palmitic, stearic, linoleic, and arachidic acids than those
from the untreated, raw seed.
Thin- layer chromatography showed that P. serotina seed oils
were devoid of the cyanogenic glycosides, amygdalin and prunasin
(Fig. 6). The patterns were similar for the toasted meal, defatted
toasted meal, and the residual meal after supercritical CO2 extrac-
tion (Rf = 0.15), corresponding to the amygdalin standard. The
prunasin standard under these conditions with an Rf of 0.52 was
absent from all samples.
Fig. 6. Thin layer chromatographic (TLC) separation of P. serotina seed components.
Lanes 1–3 denote whole meal, defatted meal and oil from toasted seed, respec-
tively; lanes 4 and 5 represent defatted meal and oil from supercritical CO2
extraction, respectively; lanes 6 and 7 are amygdalin and prunasin standards,
respectively.
The P. serotina seed contains about 21–33% of oil fraction that is
devoid of cyanide components. The oil is distinguished from the
other vegetable oils by its high refractive index, diene and triene
values. The P. serotina seed oil is somewhat unique because of its
significant a-eleostearic acid content. a-Eleostearic acid is effective
in suppressing growth in cancer cells, lowering the serum lipids in
mammals, and has been proposed as a chemotherapeutic agent
against breast cancer (Koba et al., 2002; Moon et al., 2010). There-
fore, the presence of a-eleostearic acid increases the potential
therapeutic significance of the P. serotina seed oil as a functional
food and nutraceutical ingredient, and a valuable health food
supplement.
Acknowledgements
We would like to thank Dr. Robert Bye, professor of the Institute
of Biology, National Autonomus University of Mexico. A voucher
specimen (AR-001) is deposited in the College of Chemistry, Na-
tional Autonomous University of Mexico, Mexico City.
This work was supported by the Consejo Nacional de Ciencia y
Technologia (CONACYT) Grant No. 266556.
References
Athukorala, Y., Mazza, G., & Oomah, B. D. (2009). Extraction, purification and
characterization of wax from flax (Linum usitatissimum) straw. European Journal
of Lipid Science and Technology, 111, 705–714.
Barthet, V. J., & Daun, J. K. (2002). An evaluation of supercritical fluid extraction as
an analytical tool to determine fat in canola, flax, solin, and mustard. Journal of
the American Oil Chemists’ Society, 79, 245–251.
Beltrán Sanahuja, A., Prats Moya, M. S., Maestre Pérez, S. E., Grané Teruel, N., &
Martín Carratalá, M. L. (2009). Classification of four almond cultivars using oil
degradation parameters based on FTIR and GC data. Journal of the American Oil
Chemists’ Society, 86, 51–58.
Bernardo-Gil, G., Oneto, C., Antunes, P., Rodrigues, M. F., & Empis, J. M. (2001).
Extraction of lipids from cherry seed oil using supercritical carbon dioxide.
European Food Research and Technology, 212, 170–174.
Beveridge, T. H. J., Girard, B., Kopp, T., & Drover, J. C. G. (2005). Yield and
composition of grape seed oils extracted by supercritical carbon dioxide and
petroleum ether: Varietal effects. Journal of Agricultural and Food Chemistry, 53,
1799–1804.
Bozan, B., & Temelli, F. (2002). Supercritical CO2 extraction of flaxseed. Journal of the
American Oil Chemists’ Society, 79, 231–235.
Cáceres, A., Fletes, L., Aguilar, L., Ramirez, O., Figueroa, L., Taracena, A. M., et al.
(1993). Plants used in Guatemala for the treatment of gastrointestinal disorders.
3. Confirmation of activity against enterobacteria of 16 plants. Journal of
Ethnopharmacology, 38, 31–38.
Comes, F., Farines, M., Aumelas, A., & Soulier, J. (1992). Fatty acids and
triacylglycerols of cherry seed oil. Journal of the American Oil Chemists’ Society,
69, 1224–1227.
Deineka, V. I., Gabruk, N. G., Deineka, L. A., & Manokhina, L. A. (2002). Triglyceride
composition of oil from stones of nine Rosaceae plants. Chemistry of Natural
Compounds, 38(5), 410–412.
Durmaz, G., Karabulut, I., Topçu, A., Asiltürk, M., & Kutlu, T. (2010). Roasting-related
changes in oxidative stability and antioxidant capacity of apricot kernel oil.
Journal of the American Oil Chemists’ Society, 87, 401–409.
Dyer, J. M., Chapital, D. C., Kuan, J.-C. W., Mullen, R. T., Turner, C., McKeon, T. A., et al.
(2002). Molecular analysis of a bifunctional fatty acid conjugase/desaturase
from tung. Implications for the evolution of plant fatty acid diversity. Plant
Physiology, 130, 2027–2038.
 Author's personal copy
990 I.A. Aguerrebere et al. / Food Chemistry 124 (2011) 983–990
Gatellier, Ph., Santé-Lhoutellier, V., Portanguen, S., & Kondjoyan, A. (2009). Use of
meat fluorescence emission as a marker of oxidation promoted by cooking.
Meat Science, 83, 651–656.
Hamedi, J., Malekzadeh, F., & Niknam, V. (2002). Improved production of
erythromycin by Saccharopolyspora erythraea by various plant oils.
Biotechnology Letters, 24, 697–700.
Hamedi, J., Malekzadeh, F., & Saghafi-nia, A. E. (2004). Enhancing of erythromycin
production by Saccharopolyspora erythraea with common and uncommon oils.
Journal of Industrial Microbiology and Biotechnology, 31, 447–456.
Horseley, S. B., & Meinwald, J. (1981). Glucose-1-benzoate and prunasin from Prunus
serotina. Phytochemistry, 20(5), 1127–1128.
Ibarra-Alvarado, C., Rojas, A., Luna, F., Rojas, J. I., Rivero-Cruz, B., & Rivero-Cruz, J. F.
(2009). Vasorelaxant constituents of the leaves of Prunus serotina ‘‘capulín”.
Revista Latinoamericana de Química, 37(2), 164–173.
International Union of Pure and Applied Chemistry (1985). Standard methods for the
analysis of oils, fats and derivatives (7th ed.). Oxford, UK: International Union of
Pure and Applied Chemistry.
Jennings, R. C., & Forti, G. (1975). Evidence for energy migration from photosystem I
to photosystem II and the effect of magnesium. Biochimica and Biophysica Acta,
376, 89–96.
Kaya, C., Kola, O., Ozer, M. S., & Altan, A. (2008). Some characteristics and fatty acids
composition of wild apricot (Prunus pseudoarmeniaca L.) kernel oil. Asian Journal
of Chemistry, 20, 2597–2602.
Koba, K., Akahoshi, A., Yamasaki, M., Tanaka, K., Yamada, K., Iwata, T., et al. (2002).
Dietary conjugated linolenic acid in relation to CLA differently modifies body fat
mass and serum and liver lipid levels in rats. Lipids, 37, 343–350.
Kovacevic, D., Dewez, D., & Popovic, R. (2007). Protochlorophyllide photo-
transformation and chlorophyll (ide) formation in barley etioplast fractions.
Photosynthetica, 45(3), 340–347.
Kyriakidis, N. B., & Skarkalis, P. (2000). Fluorescence spectra measurement of
olive oil and other vegetable oils. Journal of AOAC International, 83,
1435–1439.
Mazzio, E. A., & Soliman, K. F. A. (2009). In vitro screening for the tumoricidal
properties of international medicinal herbs. Phytotherapy Research, 23(3),
385–398. doi:10.1002/ptr.2636.
McCune, L. M., & Johns, T. (2007). Antioxidant activity relates to plant part, life form
and growing condition in some diabetes remedies. Journal of
Ethnopharmacology, 112, 461–469.
Mimuro, M., Hirayama, K., Uezono, K., Miyashita, H., & Miyachi, S. (2000). Uphill
energy transfer in a chlorophyll d-dominating oxygenic photosynthetic
prokaryote, Acaryochloris marina. Biochimica and Biophysica Acta (BBA) –
Bioenergetics, 1456, 27–34.
Moon, H.-S., Guo, D.-D., Lee, H.-G., Choi, Y.-J., Kang, J.-S., Jo, K., et al. (2010). Alpha-
eleostearic acid suppresses proliferation of MCF-7 breast cancer cells via
activation of PPARy and inhibition of ERK ½. Cancer Science, 101, 396–402.
Oomah, B. D., Busson, M., Godfrey, D. V., & Drover, J. C. G. (2002). Characteristics of
hemp (Cannabis sativa L.) seed oil. Food Chemistry, 76, 33–43.
Oomah, B. D., Dumon, D., Cardador-Martinez, A., & Godfrey, D. V. (2006).
Characteristics of Echinacea seed oil. Food Chemistry, 96, 304–312.
Oomah, B. D., Mazza, G., & Przybylski, R. (1996). Comparison of flaxseed meal lipids
extracted with different solvents. Lebensmittel-Wissenschaft und Technologie, 29,
654–658.
Padley, F. B., Gunstone, F. D., & Harwood, J. L. (1994). Occurrence and characteristics
of oils and fats. In F. D. Gunstone, J. L. Harwood, & F. B. Padley (Eds.), The lipid
handbook (2nd ed., pp. 47–223). London, England: Chapman and Hall.
Prashantha, M. A. B., Premachandra, J. K., & Amarasinghe, A. D. U. S. (2009).
Composition, physical properties and drying characteristics of seed oil of
Momordica charantia cultivated in Sri Lanka. Journal of the American Oil Chemists’
Society, 86, 27–32.
Russin, T. A., van de Voort, F. R., & Sedman, J. (2004). Novel method for rapid
monitoring of lipid oxidation by FTIR spectroscopy using disposable IR cards.
Journal of the American Oil Chemists’ Society, 81, 111–116.
SAS Institute, Inc. (1990). SAS/STAT User’s Guide, Version 6 (4th ed.). Cary, NC: SAS
Institute.
Spitzer, V. (1999). Screening analysis of unknown seed oils. Fett/Lipid, 101(1), 2–19.
Tósaki, A.,Vecsernyes, M., Fesus, L., Bak, I., Juhász, B., Papp, L., & Toth, S. (2008). The
application of the oil fraction obtained from sour cherry (Prunus cerasus) seed
kernel. World Patent, WO 2008/035129 A2.
Yamaguchi, K., Liggett, J. L., Kim, N.-C., & Baek, S. J. (2006). Anti-proliferative effect of
horehound leaf and wild cherry bark extracts on human colorectal cancer cells.
Oncology Reports, 15(1), 275–281.
Yu, L., Parry, J. W., & Zhou, K. (2006). Fruit seed oils. In F. Shahidi (Ed.), Nutraceutical
and specialty lipids and their co-products (pp. 73–90). Boca Raton, Florida: CRC
Press.
Yurchak, L., Dobberstein, R., & Heist, L. S. (2003). Sleep inducing antacid composition.
US Patent 2003/0013639 A1, January16, 2003.