Turkish Journal of Agriculture and Forestry Turk J Agric For

(2015) 39: 260-271

http://journals.tubitak.gov.tr/agriculture/
© TÜBİTAK
Research Article doi:10.3906/tar-1409-34

Effects of roasting and enzyme pretreatments on yield and quality of

cold-pressed poppy seed oils
Dilek DÜNDAR EMİR, Buket AYDENİZ, Emin YILMAZ*
Department of Food Engineering, Faculty of Engineering, Çanakkale Onsekiz Mart University, Çanakkale, Turkey

Received: 08.09.2014 Accepted: 20.11.2014 Published Online: 06.04.2015 Printed: 30.04.2015

Abstract: The aim of this study was to compare the effects of roasting and enzyme pretreatments on the yield and quality of cold-pressed
oils obtained from 3 poppy seed varieties (Ofis 3 (blue), Ofis 4 (yellow), Ofis 8 (white)). The oil recovery range was 21.89%–38.36% with
cold pressing. The oily cakes (meal) contained 42–168 g/kg oil and 113–166.3 g/kg moisture after the cold pressing. Physicochemical and
quality tests were conducted to evaluate and compare the efficacy of the pretreatments. Furthermore, fatty acid, sterol, and tocopherol
compositions were determined. Oil recovery was enhanced by enzyme treatment in Ofis 3 (28.31%) and Ofis 8 (38.36%), while roasting
enhanced oil yield in Ofis 4 (29.36%) samples. On the other hand, enzyme treatment caused some problems, like increases in free
acidity and peroxide value. The other physicochemical properties and minor components of the oils were not significantly affected by
the pretreatments. Furthermore, it was determined that the oily cakes can be a good source of protein. This study proved that higher
quality poppy seed oil can be produced with acceptable yield by applying the cold pressing technique coupled with a preroasting process.

Key words: Cold pressing, enzyme, oil quality, oil yield, poppy seed, roasting

1. Introduction coatings, and cosmetics. The remaining cake or meal is

Poppy (Papaver somniferum L.) is an annual plant a valuable feedstuff for dairy cattle (Kapoor, 1995; Azcan
belonging to the family Papaveraceae, including over et al., 2004). In a previous study, the oils from 8 different
100 genera. It grows in regions with hot summers and poppy varieties were Soxhlet-extracted and analyzed
midlevel rains. Poppy has been cultivated in the Eastern (Erinç et al., 2009). The main fatty acids were linoleic
Mediterranean region; in the archaeological findings acid, oleic acid, and palmitic acid. Gamma-tocopherol
of Sumer and Assyria, poppy drawings can be seen. The and alpha-tocopherol were also measured. The total mean
cultivation of poppy has long been in practice in Turkey sterol concentration of the oil samples was 2916.20 ppm.
and India. It was indicated that poppy has been cultivated The main sterols were β-sitosterol, campesterol, and Δ5-
in Turkey since Hittite times (Yayçep, 2005; TGB, 2011). avenasterol. In another study, seed oil content and fatty
Legal poppy cultivation is allowed under the rules of acid compositions of 18 cultivated Turkish poppy varieties
the United Nations, the main producer countries being were analyzed (Rahimi et al., 2011). The oil content of
Turkey, India, Australia, France, Spain, and Hungary. In the samples ranged from 35.38% to 47.95%, with linoleic
these countries, approximately 100,000 ha is annually acid, oleic acid, and palmitic acid as the main fatty acids
used for poppy production. Around 51% of the area used quantified.
worldwide for poppy cultivation is in Turkey. On the other The poppy oil literature lacks information about the
hand, Turkey provides only 17% of the legal morphine effects of different oil processing techniques on the oil and
production from poppy (TGB, 2011). meal yield and quality; this study would be an important
According to Kapoor (1995), neither poppy seed nor starting point for such studies. Furthermore, cold press
poppy seed oil contains alkaloids. Hence, both can be used oil production is a technique preferred for edible oils of
safely as human food. Poppy seeds contain around 45%– unique quality under environmentally friendly processing
54% oil and 20%–30% protein. Poppy seeds are usually used conditions. Hence, the objectives of this study were to
in bakery products for flavoring and decorative purposes. evaluate the effects of roasting and enzyme treatment
Poppy seed oil is used as an edible cooking oil locally against a control group, as preprocesses applied before
in Turkey, and is also used in the production of paints, cold pressing oil extraction technique, on the oil yield and
* Correspondence: [email protected]
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quality in 3 different poppy seed varieties. In addition, contents were constantly measured during the treatment
some properties of the cold press oily cakes (meal) were techniques and all treatment groups were set to 12%
measured in order to evaluate the quality of the resulting moisture content by water conditioning prior to cold
byproducts. Therefore, possible industrial applications to pressing. Water conditioning of the seeds was done by
produce optimum quality edible poppy seed oil by cold placing the seeds in plastic bottles, spraying the calculated
pressing technique can be developed. amount of water onto the seeds, hermetically covering the
bottles, and storing the samples at room temperature for
2. Materials and methods 24 h. The amount of water required for conditioning was
2.1. Materials calculated with the following formula: W = [(A / B) × C]
In this study, 3 different poppy seed varieties registered – C, where W indicates the amount of added water, A the
by the Turkish Grain Board (Turkish abbreviation: TMO) dry matter content of the seeds (%), B the required dry
were used. The poppy seeds of Ofis 8 (white) produced matter content (%) of the seeds, and C the amount of the
in Ulubey/Uşak, Ofis 4 (yellow) produced in Şuhut/ seeds (g).
Afyonkarahisar, and Ofis 3 (blue) produced in Ulubey/ Roasting of the poppy seeds was carried out in a Luxell
Uşak were collected from local growers. All seeds were Lx3530 type oven (Kumtel, Turkey; 1450 W) at 150 °C for
cultivated in the 2011 harvest season and were dried and 30 min. Selection of roasting temperature and duration
cleaned of foreign materials. The seeds were stored in a cool were based on literature suggestions (Şimşek, 2009) and
and dry storage area in cotton cloth bags (25 kg) during our preexperiments. The seeds were placed in metal plates
with a height of 2 cm. When the temperature in the middle
the study. Ferzim hemicellulase (60,000 U/g, optimal
point of the seed batch reached 150 °C, it was mixed up
activity at 55–65 °C and 4.0–6.5 pH) and Alphamalt BK
every 10 min for constant heat transfer. At the end of
Quick protease (12 U/g, optimal activity at 40–65 °C and
30 min of roasting, the seeds were left to cool to room
4.5–6.0 pH) were bought from local chemical suppliers.
temperature and then the moisture levels were measured.
All other chemicals and standards were of analytical grade
Water conditioning was applied to set the moisture content.
and purchased either from Merck (Germany) or Sigma-
Enzyme treatments of the poppy seeds were performed
Aldrich (USA).
by incubation with enzyme solutions. The conditions for
2.2. Analyses of the poppy seeds the enzyme treatment were based on preliminary work in
The moisture content (%) of the samples was measured the laboratory. For this purpose, 2.5 g of hemicellulase (60
with an OHAUS MB45 moisture analyzer (Switzerland) at U/g seed) was dissolved in 500 mL of 0.1 M Na2PO4 + 0.1
107 °C with 1 g sample for a 30-min drying program, while M citric acid buffer solution (6.0 pH). Due to the very small
water activity of the seeds was determined with an AQUA size of the poppy seeds, the enzyme solution was mixed
Lab 4TE instrument (Decagon Devices, USA) at room with the seeds (2.5 kg) without crushing, and the mixture
temperature according to the manual. The total amount was incubated at 60 °C for 3 h in an incubator. Then 2.5 g
of fat in the seeds was measured by the Soxhlet technique of protease (0.012 U/g seed) was dissolved in 100 mL of
according to the AOAC 920.39 method (AOAC, 2002). the same buffer, added to the seed slurry, and incubated
The total ash of the seeds was measured by the AOCS for an additional 1 h. After incubation, the enzymes were
Ba 5a-49 technique and crude protein was evaluated by inactivated by heating the seeds up to 100 °C and leaving
applying the Kjeldahl technique of AOCS Aa 5-38 (AOCS, the mixtures for 2 h at the same temperature. During the
1984). Additionally, seed color was measured with a heat treatment, water vaporized; however, roasting did not
Minolta CR300 colorimeter (Japan). A digital caliper (CD- occur. Meanwhile, the seed moisture levels were monitored
15CP, Mitutoyo Ltd., UK) was used to measure seed size. and when moisture decreased down to 12%, the batches
Hundreds of seeds were counted and weighed 3 times per were cooled down to room temperature.
each variety to determine 1000 seeds’ average weight. 2.4. Cold pressing of the poppy seeds
2.3. Preparation of the poppy seeds for cold pressing Cold pressing was performed with a laboratory scale (12
Each poppy seed variety was portioned as 2 control, 2 kg seed/h capacity) cold press machine (Koçmaksan ESM
roasting, and 2 enzyme treatment groups so that there 3710) that is a single head expeller type (kms10) with a
would be 2 equal portions for each treatment group for 1.5-kW powered engine and 0.6 kW of heating resistance.
every variety. Thus, cold pressing was applied in duplicate. The 10-mm exit die, 20-rpm screw rotation speed, and 40
Before cold pressing, the pretreatments were completed °C exit temperature were selected for all oil productions
for every portion in each treatment group and variety. as constant parameters. When oil (liquid phase) and oily
All poppy seeds were tempered to 12% moisture prior cakes (solid phase) were collected and weighed, the oily
to cold pressing with the lab scale cold press machine phase was filtered immediately through a 40-µm screen to
(Koçmaksan ESM 3710, Turkey). In this way, the moisture separate suspended materials. They were then placed into

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amber-colored and capped glasses, flushed with nitrogen, Tl 1a-64, and the amounts of unsaponifiable matter were
and kept in a fridge until analysis. The sediment levels (%) measured by ISO 3596 method (AOCS, 1984; TSE, 2002).
of the oil samples were measured. For this purpose, 5 mL The total phenolic compounds were extracted from the oil
of oil was filtered through Whatman 41 filter paper that samples twice by a water:methanol (60:40 v/v) mixture,
was previously weighed and at the end of filtration the and the total phenolic contents of the extracts were
filter paper was washed twice with 10 mL of hexane and determined by the Folin–Ciocalteu technique according
dried at 70 °C for 1 h. It was then washed 3 times with to Papoti and Tsimidou (2009) with an Agilent 8453 UV-
15 mL of diethyl ether and dried at room temperature Vis spectrophotometer (Germany). The total phenolic
overnight. Finally the filter paper was weighed, and the oil contents were calculated as mg gallic acid equivalents per
sediment levels were calculated by the following formula: 100 g oil. The antioxidant capacities of the oil samples were
level of sediment (%) = (A – B) / C × 100, where A is the evaluated in the same phenolic extracts and were expressed
final and B is the initial weight of the filter paper, and C is as Trolox equivalent antioxidant capacity (TEAC) (mmol
the amount of oil filtered. Trolox equivalents per g oil) (Re et al., 1999).
Because of the fact that the meal produced in cold 2.8. Component analyses of the poppy seed oils
pressing systems contains higher amounts of oil than that The tocopherol compositions of the samples were analyzed
obtained from industrial hot presses, the term “oily cake” in accordance with the following ISO 9936 method by an
is preferred. The oily cakes obtained through cold pressing HPLC system (Shimadzu, Japan) equipped with an LC-
were in the form of rods of 10–20 cm. Immediately after 20AT pump system, 5 µm LiCrosorb Si60, 250 × 4.6 mm
cold pressing, the resulting oily cakes were crushed with i.d. column (Hichrom, UK), and Shimadzu RT-10A-XL
a Warring blender (7011S, Warring Laboratory, USA), florescent detector (ISO, 2006). The mobile phase was
placed into zipped refrigerator bags, labeled, and frozen at 3.6% (v/v) THF in n-heptane with 1 mL min–1 flow rate at
–20 °C until analysis. 25 °C constant column temperature. Excitation at 270 nm
2.5. Analyses of the poppy seed oily cakes and emission at 310 nm were used and quantification was
The moisture content (%) of the cakes was measured with done with external standards of α-, γ-, and β-tocopherols.
an OHAUS MB45 moisture analyzer (1 g sample, 107 °C, The sterol compositions of the oil samples were
and 30 min), while fat content was measured by a Soxhlet measured by ISO 12228 method (TSE, 1999). The
apparatus according to AOAC 920.39, crude protein samples were prepared and injected by an autosampler
according to AOCS Aa 5-38, and total ash according to into a PerkinElmer Auto System XL gas chromatograph
AOCS Ba 5a-49 (AOCS, 1984; AOAC, 2002). equipped with an FID and an HP-5 (30 m × 0.32 mm ×
0.25 µm) column. Hydrogen was used as the carrier gas
2.6. Physical analyses of the poppy seed oils
at a flow rate of 30 cm3/s with a 1:50 injector split and
The density of the oil samples was measured according
injection volume was 0.2 µL. The injector and the detector
to AOCS method Cc 10c-95 and refractive indices were
temperatures were 280 and 300 °C, respectively. The
determined with an Abbe 5 (Bellingham-Stanley, UK)
column oven temperature program was set as follows:
refractometer at 20 °C (AOCS, 1984). Oil viscosity was
initial temperature was 240 °C for 0.5 min, it increased
measured with a Brookfield viscometer (model DV-II + at 5 °C min–1 to 255 °C and was held for 4 min, then it
Pro with Rheocalc software, Brookfield Eng., USA) using increased at 5 °C min–1 to 310 °C and was held for 30
an LV-SC4-18 spindle and a special sample holder. The min. GC control, data collection, and integration were
viscosity values were determined as centipoises (cP) at 20 performed by Total Chrom Navigator version 6.3.1. The
°C with 30 rpm shear rate applied. The turbidity values were phytosterols were characterized by comparison of their
measured at 20 °C using a Hach 2100-AN Turbidimeter retention times (relative to 5α-cholestane) with those of
(USA), while the instrumental color values (L, a*, b*) of commercially available standards.
the samples were measured with a Minolta Colorimeter The quantification of fatty acid methyl esters was done
CR-200 (Minolta Camera Co.). The temperature of oil using a gas chromatograph (Finnigan Trace Ultra, Italy)
samples was set to 20 °C in a water bath, and immediately equipped with an HP 88 capillary column (100 m × 0.25 mm
after taking out the samples, they were read with the i.d. with 0.2 µm film thickness; Agilent Technologies, Inc.,
instruments in the laboratory at the ambient temperature. USA). The qualification of the fatty acids was performed
2.7. Chemical analyses of the poppy seed oils with a mass spectrophotometer (Finnigan Trace DSQ,
The free fatty acid (FFA) and peroxide values of the oil USA) at 200 °C direct capillary interface temperature,
samples were determined according to AOCS methods 70 eV ionization energy level, 50–500 amu mass range
Ca 5a-40 and Cd 8-53, respectively (AOCS, 1984). with 500 amu s–1 scanning rate. A 37-component FAME
Similarly, iodine numbers and saponification numbers mixture (C4-C24, Supelco, USA) and CLA standards (Nu-
were evaluated by following AOCS methods Cd 1-25 and Check, USA) were used for fatty acid determination.

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2.9. Statistical analysis content was higher in the Ofis 8 sample. Although there
All physical, chemical, and instrumental analyses in this were some statistical differences among the water activity
study were performed twice for all varieties and treatment and moisture and ash contents of the seeds, the differences
groups. The comparison of the treatments and poppy were not large. Özcan and Atalay (2006) reported some of
seed varieties was accomplished by 2-way ANOVA and the physical and chemical properties of 7 different poppy
Tukey’s tests. Nonmetric multidimensional scaling (MDS) seed varieties. Thousand-seed weight values were between
was used to observe the cumulative relativeness of the 0.290 and 0.429 g, moisture contents were 3.39%–4.76%,
data by poppy varieties and treatments. MDS maps were amounts of crude protein were 11.94%–13.58%, crude
calculated over Z-score standardized values. All statistical ash amounts were 4.92%–6.25%, crude fiber values were
analyses were done by using Minitab 16.1.1 and SPSS 22.63%–30.08%, HCl-insoluble ash measurements were
package programs. 0.72%–1.685%, crude oil quantities were 32.43%–45.52%,
and crude energy values were 6367–6740.5 kcal/100 g. The
3. Results and discussion protein contents of the poppy seed varieties analyzed in
3.1. General properties of the poppy seeds our study were not in good agreement with this study. This
The physical and chemical properties of the 3 poppy might be due to the different analysis techniques used. On
seed varieties are shown in Table 1. There are statistically the other hand, Azcan et al. (2004) reported total protein
significant differences among the seeds. contents in yellow, white, and blue seeds as 21.8%, 21.9%,
The highest Soxhlet-extracted oil content (46.3%) was and 22.7%, respectively, and these findings are relatively
found in the blue seeds (Ofis 3), while white seeds (Ofis 8) closer to our results. To the best of our knowledge, no
contained the lowest amount (36.07%) of oil. In a previous study reporting the instrumental color values of poppy
study (Azcan et al., 2004), contrary to our results, the oil seeds is present in the literature. The color components of
contents in yellow seeds were much higher than in blue luminosity (L value) (L = 0 black, L = 100 white), a* values
and white poppy seeds, but the varieties were not stated in (+a* = red, –a* = green), and b* values (+b* = yellow, –b*
that study. Furthermore, in the same study, the mean seed = blue) of the seed samples measured according to the CIE
moisture level was reported to be 6.4%. In another study system (Pomeranz and Meloan, 1994) are reported in Table
(Erinç et al., 2009), the oil content was highest in Ofis 96 1. Bajpai et al. (1999) reported poppy seed size between 0.5
yellow seeds, and lowest in third-class white seeds. Rahimi et and 1.3 mm, and this range is similar to the findings in our
al. (2011) analyzed 18 poppy seed varieties grown in Turkey study.
and found that the highest amount of oil was in Afyon95 3.2. Oil yield and sediment contents of the poppy seeds
yellow seed (47.95%), and the lowest was in TMO2 gray The amounts of oil obtained from the 3 poppy seed
seed (35.38%). The results of our study are in accordance varieties with the treatments applied before cold pressing
with the literature. Different ranges might be due to variety, and the oil sediments measured in the oil samples (%) are
region of cultivation, and climatic factors. The protein presented in Table 2.

Table 1. Proximate analysis and seed characteristics of 3 poppy seed varieties.

Property Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Oil (g/kg) (P = 0.013) 463.01 ± 1.60 A
389.12 ± 1.17 B
360.7 ± 0.50 C
Moisture (g/kg) (P = 0.000) 64.01 ± 1.20 A 53.05 ± 0.20 B 54.20 ± 0.50 B
Ash (g/kg) (P = 0.001) 62.05 ± 0.50 A 56.17 ± 0.10 B 61 ± 0.40 A
Protein (g/kg) (P = 0.020) 193.20 ± 1.40 C 201.24 ± 3.10 B 214.01 ± 2.70 A
Water activity (aw) (P = 0.001) 0.6031 ± 0.0025 A 0.5775 ± 0.0007 C 0.5946 ± 0.0035 B
Color L (P = 0.001) 44.98 ± 0.49 C 47.61 ± 0.07 B 64.28 ± 0.51 A
a* (P = 0.000) 2.97 ± 0.17 C 7.80 ± 0.10 A 4.04 ± 0.14 B
b* (P = 0.004) –3.44 ± 0.18 C 11.94 ± 0.33 B 14.36 ± 0.20 A
Seed size (mm) (P = 0.038) 1.71 ± 0.29 B 1.16 ± 0.16 C 2.22 ± 0.30 A
1000-seed weight (g) (P = 0.000) 0.36 ± 0.06 B 0.33 ± 0.05 C 0.38 ± 0.01 A

Means followed by the same letter in the same row are not significantly different (P > 0.05).
A, B

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Table 2. The oil yield and sediment contents of the poppy seed oils after cold pressing.

Property Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Control 21.89 ± 6.89 B
27.85 ± 5.76 A
28.44 ± 4.03 B
Oil yield (%) Roasted 22.18 ± 0.28 B 29.36 ± 2.62 A 24.33 ± 7.42 B
Enzyme 28.31 ± 1.70 A 27.82 ± 1.77 B 38.36 ± 0.58 A
Control 1.92 ± 0.08 A 1.17 ± 0.31 A 2.19 ± 0.82 A
Oil sediment (%) Roasted 1.91 ± 0.07 A 1.53 ± 0.37 A 1.33 ± 0.81 A
Enzyme 1.89 ± 0.42 A 1.35 ± 0.18 A 1.68 ± 0.37 A

A, B
Means followed by the same letter in the same row are not significantly different (P > 0.05).

While the effects of seed variety (P = 0.083) and 0.492, respectively). The oil sediment measurements give
treatment (P = 0.066) were significant, their interactions (P an idea about any residual microparticles present in the
= 0.230) were not statistically significant for the amount of oil after filtration. The values ranged between 1.17% and
crude oil recovered by cold pressing. Compared to solvent- 2.19%, which is usually evaluated as a small rate and can
extracted oil content (36.07%–46.30%) shown in Table 1, be reduced by filtration or centrifugation. Such amounts
the oil recovery rates (21.89%–38.36%) were much lower of sediment did not create any visual turbidity problems
in all treatments by cold pressing (Table 2). The enzyme in the samples. As in virgin olive oil production, natural
treatment in Ofis 3 and Ofis 8 samples, and roasting in Ofis decantation in stainless steel storage tanks can be easily
4 samples, yielded more oil than the control, indicating applied to reduce any turbidity problems.
some limited positive effects of the treatments on the oil 3.3. Analyses of the poppy seed oily cakes
yield in cold pressing. Hence, these pretreatments can Some properties of the oily cakes of the poppy seed
be suggested for application prior to cold pressing, but varieties analyzed within this study are given in Table 3.
the quality parameters of the resulting oils must also be The moisture contents ranged between 113.4 and 166.3
evaluated. Soto et al. (2007) showed that application of g/kg. Although seed moisture level was adjusted to 12%
Olivex and Celluclast enzymes to borage seeds before cold by water conditioning before cold pressing, the oily cakes
pressing enhanced oil yield significantly and did not affect had higher moisture levels. This result might be due to the
oil quality. Erinç et al. (2009) reported that the Soxhlet- proportional change after oil exclusion. While the effect of
extracted oil contents of 8 poppy varieties ranged from 483 treatment (P = 0.021) on cake moisture was significant, the
g/kg to 527 g/kg. In another study, the solvent-extracted effects of seed variety (P = 0.352) and variety–treatment
oil contents of different poppy varieties were between 35% (P = 0.978) interactions were not statistically significant.
and 48% (Rahimi et al., 2011). As can be seen from these The crude protein contents of the cakes, measured by the
results, the oil yield obtained through solvent extraction is Kjeldahl technique, were determined to vary between 210
distinctly higher than that of cold pressing. On the other and 280 g/kg, so there is a proportional increase in the
hand, cold-pressed oil can be utilized without refining, protein amount of cakes compared to that of the seeds.
and hence most of the minor components remain in the Similarly, the effects of seed variety, treatment, and their
oil. Furthermore, the oil retained in the oily cake can be interactions were unimportant on cake protein level. On
readily extracted with a solvent, if desired. It might be the other hand, seed variety and treatment interactions (P
suggested from our experiments that oil yield in cold = 0.005) significantly affected the remaining oil content
pressing can be enhanced by increasing press heat and of the samples, while variety and treatment were not
decreasing seed moisture content or by prolonged enzyme significant alone. After cold pressing, there was 42–170
incubation, but the resulting oil becomes dark, burned, g/kg oil remaining in the cakes. The total ash contents of
and sensorially unacceptable. Hence, if cold-pressed oil the cakes ranged between 51 and 99 g/kg. The Turkish
will be utilized without a refining process as virgin oil, Standard TS 319 named “Poppy seed meal (cake)” provides
the oil yield might be increased through optimization of some values about expeller- and solvent-extracted poppy
processing conditions to a limited degree. seed cakes (TSE, 2003). The standard indicates that poppy
The effects of seed variety, treatments, and their seed cakes should contain 30%–35% protein, 17% crude
interactions on the oil sediment content (%) were not cellulose, 3%–6% oil, 9% ash, 1% foreign matter, and 12%
statistically significant (P = 0.159, P = 0.808, and P = moisture. Since a cold pressing system was used in this

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Table 3. Proximate analyses of the poppy oily cakes (meals) after cold pressing.

Property Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Control 113.40 ± 31.70 125.70 ± 4.80 144.50 ± 13.20
Roasted 142.20 ± 10.30 126.10 ± 15.20 134.12 ± 5.70
Moisture (g/kg)
Enzyme 166.30 ± 31.30 159.10 ± 8.30 164.10 ± 13.40
Control 250.00 ± 10.01 225.00 ± 7.00 245.00 ± 21.20
Protein (g/kg)
Roasted 235.00 ± 35.00 250.00 ± 10.10 255.10 ± 21.20
(N × 6.25)
Enzyme 210.00 ± 28.30 280.00 ± 10.10 265.00 ± 7.00
Control 60.10 ± 7.70 Bb 155.20 ± 34.10 Aa 148.30 ± 10.00 Aa
Oil (g/kg) Roasted 108.20 ± 1.14 Aab 138.60 ± 30.20 Aa 132.70 ± 13.00 Aa
Enzyme 168.60 ± 34.80 Aa
42.10 ± 6.50 Bb
100.3 ± 31.60 ABa
Control 94.01 ± 4.20 Aa 51.50 ± 8.50 Bb 90.50 ± 9.50 Aa
Ash (g/kg) Roasted 77.50 ± 18.50 Aa 82.50 ± 2.50 Aab 87.00 ± 4.00 Aa
Enzyme 83.00 ± 10.50 Aa
99.50 ± 7.00 Aa
76.50 ± 13.40 Aa

Means followed by the same letter in the same row are not significantly different (P > 0.05).
A, B

Means followed by the same letter in the same column are not significantly different (P > 0.05).
a, b

study, the values are different. Although the oil recovery study provides simple but important data for poppy seed
rate is lower in cold pressing systems, the produced oil is oil characteristics.
solvent-free, of high quality, and edible. Moreover, oily The FFA levels of the oil samples ranged from 1.3% to
cakes from cold pressing can be valuable feedstock and can 6.9% and there was no statistically significant difference
be utilized for human food production, since they are free between varieties (P = 0.285), treatments (P = 0.173), and
of solvents and chemicals. their interactions (P = 0.997). Although roasting reduced
3.4. Physicochemical characteristics of the poppy seed the FFA level a little, enzyme treatment caused some
oils minor increases compared to the control. In general, the
Some physicochemical analyses were applied to the poppy FFA level should not exceed 3% in virgin oils. Otherwise,
seed oil samples produced in this study and the results are neutralization is required to reduce the acidity because
shown in Table 4. acidic oils are sensorially unacceptable due to sour taste
The refractive indices of the samples ranged between and burning sensation (Kayahan and Tekin, 2006). Cold
1.4748 and 1.4764, and there were no significant pressing can yield edible and very high quality oil, though
differences among the varieties and treatments. A similar seed roasting seems to be a prerequisite treatment for
trend was evident for the viscosity (42.8–44.3 cP) and both better yield and higher quality. Enzyme treatment
specific gravity (0.91947–0.92072 g/cm3) values. The can enhance oil yield (Table 2) but oil quality is reduced,
turbidity of the samples presented a rather larger variation possibly due to the lipolytic activity present as impurities in
(1–181 NTU) and significant differences. The highest the enzymes used, or due to oil hydrolysis that developed
turbidity values were observed in the control groups and during incubation of seeds in an aqueous enzyme solution.
Ofis 8 samples, though roasting and enzyme treatment Therefore, it can be suggested that enzyme treatment
were found to reduce turbidity significantly. Hence, it can would be better if the oil is going to be neutralized
be claimed that to produce more clear cold-pressed edible before consumption. The peroxide values of the samples
oil, roasting can be a valuable pretreatment application. ranged from 0.68 to 2.98 mEq O2/kg oil. The values
There were no important differences among the L and were dependent on variety (P = 0.017) and treatment (P
b* color components of the samples, while the a* values = 0.029), but were not dependent to variety–treatment
exhibited treatment-dependent variation. Azcan et al. interactions (P = 0.149). The peroxide values were also
(2004) reported that the mean refractive index was 1.4709 significantly higher in enzyme-treated samples than in the
and mean specific gravity was 0.940 g/cm3 for poppy seed roasted and control groups. It is quite possible that during
oil. These values and those measured in this study are the incubation period in the enzyme solution, air-based
in accordance. There was no report in the literature for oxidation may have taken place. Here again, if higher
viscosity, turbidity, and color values to compare, so our yield is the aim of enzyme treatment, neutralization may

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Table 4. The physicochemical properties of the poppy seed oils.

Property Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Control 1.475 ± 0.001 1.475 ± 0.002 1.475 ± 0.001
Refractive index (20 °C) Roasted 1.475 ± 0.005 1.476 ± 0.001 1.475 ± 0.002
Enzyme 1.475 ± 0.002 1.474 ± 0.001 1.476 ± 0.001
Control 43.20 ± 0.10 43.10 ± 0.80 43.8 ± 1.10
Roasted 43.90 ± 0.30 44.30 ± 0.70 43.3 ± 0.20
Viscosity (20 °C, cP)
Enzyme 42.80 ± 0.30 43.60 ± 0.60 43.2 ± 0.40
Control 0.9202 ± 0.0064 0.9205 ± 0.0003 0.9194 ± 0.0007
Specific gravity
Roasted 0.9201 ± 0.0009 0.9200 ± 0.0001 0.9198 ± 0.0009
(20 °C, g/cm3)
Enzyme 0.9207 ± 0.0019 0.9203 ± 0.0005 0.9202 ± 0.0004
Control 43.00 ± 0.00 Ca 168.00 ± 1.00 Ba 181.50 ± 1.50 Aa
Turbidity (20 °C, NTU) Roasted 1.00 ± 0.00 Ab 1.50 ± 0.50 Ab 1.00 ± 0.00 Ab
Enzyme 1.00 ± 0.00 Ab
1.25 ± 0.50 Ab
3.50 ± 0.50 Ab
Control 54.09 ± 0.05 52.42 ± 1.02 52.53 ± 0.92
Color L Roasted 54.05 ± 0.23 54.02 ± 0.37 53.31 ± 0.35
Enzyme 53.64 ± 0.19 54.68 ± 0.05 52.08 ± 0.92
Control 0.63 ± 0.09 Aa 1.32 ± 0.40 Aa 1.87 ± 0.02 Ab
a* Roasted 0.31 ± 0.24 Aa 0.09 ± 0.05 Aa 1.28 ± 0.01 Ab
Enzyme 1.29 ± 0.41 Ba 0.24 ± 0.19 Ba 3.31 ± 0.83 Aa

  Control 4.59 ± 0.03 1.75 ± 1.48 3.65 ± 1.28

b* Roasted 4.84 ± 0.03 4.87 ± 0.17 4.73 ± 0.20
  Enzyme 4.76 ± 0.55 4.54 ± 0.04 3.27 ± 0.99
Control 4.01 ± 1.50 3.40 ± 0.90 6.40 ± 1.72
Free fatty acid
Roasted 1.50 ± 0.10 1.30 ± 0.10 3.20 ± 0.31
(% linoleic acid)
Enzyme 4.10 ± 2.30 4.50 ± 3.40 6.90 ± 3.50
Control 0.68 ± 0.44 0.95 ± 0.24 1.37 ± 0.28
Peroxide value Roasted 0.82 ± 0.11 0.84 ± 0.11 1.29 ± 0.06
(mEq O2/kg oil) Enzyme 1.82 ± 0.39 0.72 ± 0.23 2.98 ± 0.78
Control 132.50 ± 5.0 133.50 ± 15 137.01 ± 20
Roasted 141.10 ± 60 135.01 ± 10 135.05 ± 40
Iodine number (g/100 g oil)
Enzyme 134.10 ± 10 137.50 ± 25 138.50 ± 45
Control 178.61 ± 5.28 179.31 ± 3.98 184.28 ± 0.05
Saponification number Roasted 186.33 ± 1.99 183.49 ± 0.59 187.63 ± 3.50
(mg KOH/g oil) Enzyme 183.77 ± 1.11 185.79 ± 0.87 182.79 ± 2.12
Control 1.02 ± 0.02 Aa
0.91 ± 0.08 Aa
0.67 ± 0.05 Bb
Roasted 0.65 ± 0.04 Bb
0.85 ± 0.05 Aab
0.79 ± 0.11 ABab
Unsaponifiable matter (%)
Enzyme 0.84 ± 0.01 ABab 0.69 ± 0.01 Bb 0.89 ± 0.03 Aa
Control 2.63 ± 0.28 Aa
1.69 ± 0.05 Ab
1.33 ± 0.16 Ab
Total phenolics Roasted 2.48 ± 0.49 Aa 3.47 ± 0.28 Ab 2.42 ± 0.14 Aab
(mg GA/100 g oil) Enzyme 1.01 ± 0.74 Ca
7.59 ± 0.81 Aa
4.28 ± 0.76 Ba
Control 31.23 ± 3.20 22.61 ± 1.75 30.37 ± 0.54
Roasted 23.25 ± 3.58 20.26 ± 1.56 27.14 ± 2.06
TEAC (mmol Trolox/g oil)
Enzyme 32.03 ± 2.14 26.62 ± 3.10 43.90 ± 15.3

A, B
Means followed by the same letter in the same row are not significantly different (P > 0.05).
a, b
Means followed by the same letter in the same column are not significantly different (P > 0.05).

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be required. Depending on the purpose of cold pressing, 3.5. Compositional characteristics of the poppy seed oils
a decision of whether or not to apply enzyme treatment The tocopherol composition of the oil samples is presented
could be made. The iodine numbers of the samples in Table 5.
varied between 132.50 and 141.00 g/100 g oil, and none The levels of γ-tocopherol were affected by both
of the factors were found to be statistically significant. A variety (P = 0.001) and treatment (P = 0.034), but not by
similar case was evident for the saponification numbers their interactions (P = 0.694). Contrarily, α-tocopherol
of the samples (178.61–187.63). Unsaponifiable matter levels were affected by variety (P = 0.001), treatment (P
contents were between 0.65% and 1.02%, and only the = 0.027), and their interactions (P = 0.042). Erinç et al.
effect of variety and treatment interaction was statistically (2009) reported g-tocopherol levels of 195.37–280.85 ppm
significant (P = 0.002). Firestone (1999) reported iodine and α-tocopherol levels of 21.99–45.83 ppm in poppy seed
numbers of 132–146, saponification numbers of 188– oil samples. Another study (Bozan and Temelli, 2008)
196, and unsaponifiable matter content of 0.4%–1.2% reported total tocopherol content as 11.0 mg/100 g poppy
for poppy seed oil. The findings of our study are in good seed oil, and the findings of our study are in agreement
agreement with the literature. On the other hand, Azcan et with the literature. In general, it can be observed from
al. (2004) reported a mean saponification number of 234, Table 5 that enzyme treatment can reduce tocopherol
unsaponifiable matter content of 1.03%, and a peroxide content by a small amount.
value of 39 for Turkish poppy seed oil samples. As can be The sterol compositions of the samples were also
seen from Table 4, the reported peroxide value is very high analyzed and the results are given in Table 6.
compared to our findings, because freshness level of the It was observed that most of the sterol components
seeds as well as the oil storage conditions can be different. were affected by both seed variety and treatment type.
The total phenolic contents and antioxidant capacities From Table 6, it is obvious that neither enzyme treatment
(TEAC) of the samples were in the ranges of 1.01 to 7.59 nor roasting influenced sterol composition in a definite
mg GA/100 g oil and 20.26 to 43.91 mmol Trolox/g oil, pattern. Of course there were some changes compared
respectively. The phenolic contents were significantly to the control group, but it is difficult to conclude that
affected by variety (P = 0.001), treatment (P = 0.001), and treatment had a definite negative or positive effect on the
their interactions (P = 0.000), whereas the differences sterol compositions of the samples. Erinç et al. (2009)
in the TEAC values were not statistically significant. No reported a mean total sterol content of 2916.20 ppm in
report about the total phenolic content and antioxidant 8 different seed varieties, with main sterol components
capacity of poppy seed oils is present in literature. Another being β-sitosterol (663.91–3244.39 ppm), campesterol
cold-pressed oil, namely virgin olive oil, is a good example (228.5–736.50 ppm), and Δ5-avenasterol (103.90–425.02
for data about these parameters. Öğütçü and Yılmaz (2009) ppm). In another source, the sterol composition of poppy
reported that total phenolics of 30–208 mg GA/kg oil and seed oil was given as: β-sitosterol 68%, campesterol 22%,
TEAC values of 0.60–5.61 Trolox Eq/kg oil were present in stigmasterol 3%, and either Δ5- or Δ7-avenasterol 2%
olive oils. Hence, our study provides the first data on these (Firestone, 1999). In general, the sterol data of our study
parameters for cold-pressed poppy seed oils. are similar to the reported data in the literature. It can be

Table 5. The tocopherol composition of the poppy seed oils.

Tocopherol Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Control 22.90 ± 0.20 33.5 ± 2.40 22.80 ± 1.30
Roasted 24.50 ± 0.40 33.90 ± 4.60 19.70 ± 0.50
Alpha-tocopherol (mg/kg)
Enzyme 20.90 ± 2.20 26.90 ± 4.30 13.20 ± 3.20
Control 279.00 ± 18.40 Aa 247.80 ± 5.70 Aa 249.8 ± 11.30 Aa
Gamma-tocopherol (mg/kg) Roasted 278.50 ± 5.10 Aa 241.40 ± 15.60 Aa 227.40 ± 9.40 Aa
Enzyme 267.40 ± 12.40 Aa 246.10 ± 9.60 Aa 156.10 ± 23.90 Bb
Control 301.90 ± 25.80 281.20 ± 11.40 272.60 ± 14.20
Total tocopherol (mg/kg) Roasted 302.90 ± 1.30 275.20 ± 28.40 247.10 ± 12.60
Enzyme 288.30 ± 20.70 272.90 ± 19.60 169.20 ± 38.30

Means followed by the same letter in the same row are not significantly different (P > 0.05).
A, B

Means followed by the same letter in the same column are not significantly different (P > 0.05).
a, b

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Table 6. The sterol composition of the poppy seed oils.

Sterol (ppm) Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)

Control 1.28 ± 0.05 Bb
3.50 ± 0.04 Aa
4.02 ± 0.25 Aa
Cholesterol Roasted 3.06 ± 1.15 ABab
1.44 ± 0.26 Bb
4.46 ± 0.85 Aa
Enzyme 3.44 ± 0.21 Aa 3.22 ± 0.17 Aab 2.65 ± 0.01 Aa
Control 1.81 ± 0.38 nd nd
Brassicasterol Roasted nd 2.61 ± 0.32 nd
Enzyme nd nd nd
Control 35.14 ± 0.97 Bb 59.82 ± 2.73 Aa 55.68 ± 0.21 Aa
24-Methylene cholesterol Roasted 42.31 ± 0.34 Aab 30.77 ± 2.28 Bb 35.78 ± 2.96 ABc
Enzyme 45.61 ± 2.57 Ba
63.42 ± 0.64 Aa
48.31 ± 0.34 Bb
Control 280.79 ± 2.78 Bc
469.67 ± 2.06 Aa
466.7 ± 10.7 Aa
Campesterol Roasted 403.02 ± 1.03 Ba 266.21 ± 0.43 Cc 428.58 ± 6.41 Ab
Enzyme 381.19 ± 0.87 Bb
449.99 ± 3.12 Ab
432.32 ± 0.11 Ab
Control 44.29 ± 0.75 Bc 63.88 ± 0.29 Aa 65.93 ± 1.72 Aa
Stigmasterol Roasted 54.24 ± 1.99 Ab
39.60 ± 0.16 Bc
52.51 ± 0.43 Ac
Enzyme 56.96 ± 0.21 Aa
58.75 ± 0.52 Ab
59.36 ± 0.08 Ab
Control 3.62 ± 1.31 Bb 14.61 ± 0.51 Aa 11.49 ± 2.42 Aa
Delta-7 campesterol Roasted 3.77 ± 3.77 ABb
1.77 ± 0.05 Bb
10.01 ± 0.150 Aa
Enzyme 12.37 ± 2.25 Aa 15.73 ± 0.22 Aa 9.51 ± 0.01 Aa
Control 1.90 ± 1.90 12.68 ± 1.09 nd
Delta-5,23 stigmastadienol Roasted nd nd nd
Enzyme 9.13 ± 1.64 15.26 ± 0.22 nd
Control 23.48 ± 0.51 Ba 21.73 ± 0.04 Ba 27.67 ± 1.88 Aa
Clerosterol Roasted 22.98 ± 0.34 Aa 14.20 ± 0.30 Bb 23.44 ± 0.18 Ab
Enzyme 23.57 ± 1.06 Aa
20.53 ± 0.02 Ba
23.93 ± 0.01 Ab
Control 920.70 ± 12.20 Cc
1221.80 ± 13.10 Ba
1554.70 ± 50.55 Aa
Beta-sitosterol Roasted 1321.90 ± 16.80 Aa 697.88 ± 6.99 Bb 1385.80 ± 15.40 Ab
Enzyme 1217.70 ± 11.40 Bb
1142.70 ± 26.80 Ba
1363.40 ± 0.10 Ab
Control 114.24 ± 1.47 Bb 245.86 ± 1.19 Aa 266.27 ± 9.93 Aa
Delta-5 avenasterol Roasted 240.58 ± 4.57 Aa
80.45 ± 0.23 Bb
245.14 ± 3.50 Aa
Enzyme 220.2 ± 13.60 Aa
236.73 ± 4.25 Aa
243.64 ± 0.25 Aa
Control 0.83 ± 0.06 Bb 12.80 ± 0.34 Aa 13.89 ± 0.67 Aa
Delta-5,24 stigmastadienol Roasted 11.69 ± 2.35 Aa
1.64 ± 0.08 Bb
14.55 ± 0.16 Aa
Enzyme 12.47 ± 0.38 Aa 13.60 ± 0.33 Aa 13.05 ± 0.20 Aa
Control 1.19 ± 0.07 nd nd
Delta-7 stigmastenol Roasted nd nd nd
Enzyme nd nd nd
Control 2.06 ± 0.12 Bb 16.32 ± 1.00 Aa 15.28 ± 0.88 Aa
Delta-7 avenasterol Roasted 15.82 ± 0.13 Aa 2.97 ± 0.44 Bb 16.27 ± 0.26 Aa
Enzyme 13.65 ± 2.38 Aa
16.96 ± 0.36 Aa
16.22 ± 0.48 Aa
Control 1431.30 ± 21.60 Cb
2142.60 ± 17.70 Ba
2481.60 ± 78.80 Aa
Total sterols Roasted 2119.40 ± 3.80 Aa 1139.50 ± 9.50 Bb 2216.50 ± 22.40 Ab
Enzyme 1996.30 ± 12.20 Ba
2036.90 ± 35.10 Ba
2212.40 ± 0.20 Ab

Means followed by the same letter in the same row are not significantly different (P > 0.05).
A, B

Means followed by the same letter in the same column are not significantly different (P > 0.05). nd : not dedected.
a, b

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claimed that enzyme treatment and roasting had no effects visualized simultaneously by MDS, which provides
on the sterol compositions of the poppy seed oil samples. a closeness map on dimensions. In this statistical
Table 7 includes the fatty acid compositions of the technique, the validity of the map is defined by the
samples. Generally, the levels of the detected 5 major fatty stress value; if the stress value is under 0.025, this is
acids in the oil samples were not found to be dependent on classified as very good, 0.025–0.05 as good, 0.05–0.1 as
the seed variety and the treatments or their interactions. acceptable, and 0.1–0.20 as poor validity. The closeness
Moreover, the amount of each individual fatty acid of the 15 measured parameters within each variety to the
seemed to be unaffected by either enzyme treatment or treatments (control, roasting, and enzyme treatment)
roasting. The fatty acid compositions of the samples were is presented in Figure 1a. The groups were very well
very similar to those reported in the literature. Bozan separated, with a stress value of 0.00618. In fact, the
and Temelli (2008) reported major fatty acids of poppy control samples are completely separated from the other
seed oil as 73% linoleic, 10% palmitic, and 13% oleic 2 groups, but also located far from each other. Contrarily,
acids. Azcan et al. (2004) reported GC/MS-analyzed roasted and enzyme-treated samples are well separated
fatty acid composition of poppy seed oil as stearic acid of but very closely located within the groups. These results
2.5%–3.2%, linolenic acid of 0.4%–0.6%, palmitic acid of once again indicate that the applied pretreatments caused
10%–13%, oleic acid of 16.1%–24.7%, and linoleic acid of some measurable differences among the samples. By a
56.4%–69.2%. Additionally, it was indicated that a major similar approach, the distribution of only seed varieties
fatty acid, linoleic acid, was highest (69.2%) in white seeds by the 15 measured parameters is shown in Figure 1b.
and lowest (56.4%) in blue seeds (Azcan et al., 2004). Obviously Ofis 8 (white seed) is definitely separated from
Another study reported that the fatty acid composition the other 2 varieties. The parameters of Ofis 3 (blue) and
of poppy seed oil was as follows: 12.20% palmitic, 0.27% Ofis 4 (yellow) seeds also form 2 definite separate groups,
palmitoleic, 2.30% stearic, 22.19% oleic, 59.87% linoleic, but closer to each other. Hence, this map indicates that
1.30% linolenic, 0.67% arachidonic, 0.16% gadoleic, and some differences in the measured parameters are also
0.29% erucic acid (Ryan et al., 2007). All these studies and dependent on the seed varieties.
our findings indicate that poppy seed oil is rich in linoleic Cold pressing is a versatile, easy-to-operate, cheap, and
acid, and because it includes some oleic and palmitic acid, high-quality oil production system. Enzyme treatment
it can be classified as a balanced nutritious edible oil. and roasting of oil seeds can improve oil yield, but enzyme
The relationship among the seed varieties and treatment in particular may cause some problems, like
treatment types by the measured parameters can be hydrolysis and oxidation in oil quality due to the longer

Table 7. Fatty acid composition of the poppy seed oils.

Fatty Acid (%) Treatment Ofis 3 (blue) Ofis 4 (yellow) Ofis 8 (white)
Control 9.51 ± 0.08 8.70 ± 0.09 8.90 ± 0.13
Roasted 9.40 ±0.05 8.70 ± 0.03 8.80 ± 0.01
Palmitic acid (C16:0)
Enzyme 9.40 ± 0.02 8.70 ± 0.00 8.70 ± 0.12
Control 2.30 ± 0.03 2.50 ± 0.03 2.30 ± 0.08
Roasted 2.20 ± 0.01 2.60 ± 0.01 2.20 ± 0.01
Stearic acid (C18:0)
Enzyme 2.20 ± 0.01 2.50 ± 0.00 2.30 ± 0.13
Control 13.60 ± 0.01 14.30 ± 0.05 13.60 ± 0.43
Roasted 13.50 ± 0.05 14.30 ± 0.06 13.20 ± 0.01
Oleic acid (C18:1)
Enzyme 13.60 ± 0.06 14.70 ± 0.00 13.70 ± 0.57
Control 74.20 ± 0.10 73.90 ± 0.17 74.50 ± 0.70
Roasted 74.21 ± 0.06 73.10 ± 0.31 75.20 ± 0.01
Linoleic acid (C18:2)
Enzyme 74.15 ± 0.07 73.50 ± 0.01 74.70 ± 0.55
Control 0.50 ± 0.01 0.50 ± 0.01 0.60 ± 0.00
Roasted 0.50 ± 0.00 0.60 ± 0.01 0.60 ± 0.01
Gamma-linolenic acid (C18:3)
Enzyme 0.50 ± 0.01 0.60 ± 0.00 0.60 ± 0.04

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Figure 1. Employment of statistical multidimensional scaling to determine cumulative relativeness among poppy seed varieties
(stress: 0.00618, RSQ: 0.99992) (a), and pretreatments (stress: 0.00618, RSQ: 0.99992) (b), prior to cold pressing oil processing.
Abbreviations: co8: control Ofis 8, co3: control Ofis 3, co4: control Ofis 4, ro8: roasted Ofis 8, ro3: roasted Ofis 3, ro4: roasted Ofis
4, eo8: enzyme Ofis 8, eo3: enzyme Ofis 3, eo4: enzyme Ofis 4.

incubation period, that may lead to deteriorative reactions. value. It can be concluded that high-quality edible poppy
In addition, the oily cake obtained after cold pressing was seed oils can be produced by cold pressing coupled with a
free of solvents and could be directly used as feedstock or preroasting process.
even processed as human food or food additives. The fatty
acid, tocopherol and sterol compositions, total phenolics, Acknowledgment
and antioxidant capacity values of the poppy seed oil This study was funded by TÜBİTAK (the Scientific and
samples have proved again that cold-pressed poppy seed Technological Research Council of Turkey) Project No:
oil is a safe, edible oil with high quality and nutritious 111O618. The authors are grateful for the funding.

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