488 DOI 10.1002/ejlt.200600026 Eur. J. Lipid Sci. Technol.

108 (2006) 488492

Ehsan Jenab Canola oil extracted by supercritical carbon dioxide

Karamatollah Rezaei
Zahra Emam-Djomeh and a commercial organic solvent
Department of Food Science Samples of crushed and cooked canola seeds (Okapy Double Zero) were extracted
and Engineering, using supercritical carbon dioxide (SCCO2) (34.0 MPa and 40.0 7C) and a commercial
Faculty of Agricultural Biosystem organic solvent (AW406). Oil solubility was obtained through several stepwise extrac-
Engineering, tions under the conditions of this study, and then three additional extractions were
The University of Tehran,
performed to measure fatty acid compositions, iodine values, chlorophyll concentra-
Karaj, Iran
tions and unsaponifiable matter. The yield of SCCO2 extraction was lower than that
after extraction with AW406 solvent, due to the incomplete SCCO2 extraction process.
Fatty acid composition analysis showed that the SCCO2-extracted oil was slightly
higher in polyunsaturated fatty acids and lower in erucic and behenic acids. However,
iodine values and unsaponifiable matter did not indicate significant differences
(p .0.05) in the two extracted oils. The chlorophyll concentration of SCCO2-extracted
oil was lower than that in the AW406 solvent, and as a result, the color of SCCO2-
extracted oil was lighter.

Keywords: Canola oil, extraction, quality, solubility, supercritical.

1 Introduction governments and industries in finding alternative solvents

Canola oil is obtained from genetically modified seeds of
Brassica napus and B. rapa [1]. These cultivars contain
,2% erucic acid in their oil and ,30 mg glucosinolates
per gram of dry meal. Canola seeds contain 4148 wt-%
(dry-based) oil with very low levels of saturated fatty acids
(,6%), high levels of oleic acid (,60%) and intermediate
levels of linoleic and linolenic acids (,20 and 10%,
respectively) [1]. Because of its higher oil content and
numerous agricultural advantages, canola seed is getting
worldwide attention.
For years, commercial extraction-grade hexane has been
used as a solvent for crude edible oil extraction. Operat-
Research Paper
[24]. Common gases, such as carbon dioxide, in their
supercritical state have properties and extraction capaci-
ties very similar to liquids [5].
Other than having numerous unique features of its own,
supercritical fluid extraction (SFE) is a process that also
carries many characteristics of conventional solvent
extraction. The technique is similar to conventional sol-
vent extraction in that the material to be extracted is
withdrawn from the substrate by using a solvent. But
rather than being a liquid, this solvent is a fluid above its
critical point [6].
In its supercritical state, carbon dioxide is the most com-

ing losses of solvent ranging from 0.2 to 2.0 gallons per
ton of seed processed, continuous increases in the price
of petroleum oil derivatives, occasional scarcities of hex-
ane, toxicological and environmental safety issues, high
flammability, concerns about the solvent residue left in
the extracted edible oils, non-selectivity for triacylglycer-
ols as the major components of the edible oils, co-
extraction of phospholipids and pigments with the oils,
major refining losses and enormous energy requirements
for the evaporation of the solvent contribute to the pro-
duction costs, which have augmented the interest of
Correspondence: Karamatollah Rezaei, Department of Food
Science and Engineering, Faculty of Agricultural Biosystem Engi-
neering, The University of Tehran, P.O. Box 3158778659, Karaj,
Iran. Phone: 198 261 2235124, Fax: 198 261 2248804, e-mail:
krezaee@ut.ac.ir
monly used supercritical solvent. This is due to its non-
toxicity, non-flammability, having low cost, being readily
available in bulk quantities, ease of removal from the
extracted materials and having a low critical temperature
and pressure (31.1 7C and 7.4 MPa, respectively) [710].
There are many reports in the literature studying the
extraction of oilseeds, including canola seeds, using
supercritical carbon dioxide (SCCO2) [6, 9, 11, 12], but
quality data to compare SCCO2-extracted oil with that
extracted by an organic solvent are needed to properly
evaluate the potential of SCCO2 extraction in the oilseed
industry. Therefore, in this study, certain quality char-
acteristics such as fatty acid composition, iodine value,
the concentration of chlorophyll and the amounts of
unsaponifiable matter of oils extracted by SCCO2 and a
commercial organic solvent (AW406) are compared.

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Eur. J. Lipid Sci. Technol. 108 (2006) 488492
2 Materials and methods
2.1 Preparation of seed samples
Okapy Double Zero canola seeds were prepared at the
Varamin Crude Edible Oil Extraction Factory (Varamin,
Iran). The impurities inside the bulk of oilseeds were
separated using a sieve (mesh number = 18). The oil
and moisture contents of cleaned canola seeds were
determined in triplicate using an NMR Analyzer (model
MQ 20 Minispec; Bruker Corporation, Hamburg, Ger-
many). The oil content of the canola seeds was
43.6 6 0.2 wt-% (dry-based). The canola seeds were
crushed using a Moulinex mill (Lyon, France) and then
the size distribution of the seed particles was deter-
mined using sieves of different mesh numbers (Tab. 1).
To improve the extraction efficiency, crushed canola
seeds were cooked for a short period (30 min) at 90 7C
[9, 13].
Tab. 1. Distribution of seed particle sizes.
Size range [mm] Percent of total mass
,0.5 21.0 6 1.5
0.51 52.0 6 1.2
11.4 22.1 6 0.8
.1.4 4.3 6 0.1
2.2 Organic solvent extraction
To perform the conventional oil extraction, a pilot-scale
Soxhlet extraction unit was operated for 9 h using AW406,
which is a commercial organic solvent applied to the
extraction of crude edible oils. Based on the manu-
facturer data, the n-hexane content of this solvent was
,30% (with a total hexane content of ,70%).
2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Quality comparison of crude canola oils 489
2.3 SCCO2 extraction
The equipment used for the SFE operation (Fig. 1) was
assembled in the Faculty of Chemistry (Isfahan Industrial
University, Isfahan, Iran). CO2 was cooled to prevent its
gasification and introduced into an HPLC pump with
cooled head (model PU-980; Jasco Corporation, Tokyo,
Japan). A stainless steel extraction vessel with a capacity
of ,50 mL was loaded with ,35.00 g crushed and
cooked canola seeds and placed in an oven
(40.0 6 1.0 7C). CO2 flow rate was maintained at
,1.6 6 0.2 L/min (under the expanded atmospheric con-
ditions). A backpressure regulator located before the
expansion valve was used to adjust the system pressure.
To avoid excessive freezing of the extracted oil, the
backpressure regulator was heated at 80 7C during the
extraction. The extracted oil was collected in a glass re-
ceiver at atmospheric pressure held in an ice bath. To
measure the flow rate of the expanded CO2, it was pas-
sed through a gas meter from Shinagava Corporation
(Tokyo, Japan). For determining the oil solubility in SCCO2
under the conditions of this study, two samples were
initially extracted in several fractions (until ,430 L of CO2
consumption), and then three extra extractions for 6 h
were performed to measure the extraction yield and to
collect samples for chemical composition analysis.
2.4 Methods of analysis
The yields of the two extraction methods were measured
gravimetrically using an analytical balance. To determine
the fatty acid compositions of the extracted oils, aliquots
of the oils were converted into their methyl esters
according to Rezaei et al. [14], and after extraction by n-
hexane, samples were analyzed using a gas chromatog-
raphy system (model GC Star 3400; Varian Corporation,
Palo Alto, CA, USA) equipped with a high-resolution cap-
illary column (DB-23; J&W Scientific, Folsom, CA, USA)
Fig. 1. Schematic representation of the SFE
apparatus used in this study. (1) CO2 cylinder,
(2) demoisturizer, (3) cooler, (4) HPLC pump,
(5) oven, (6) extraction cell, (7) backpressure
regulator, (8) receiver in ice bath, (9) gas meter,
(10) vent.
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490 E. Jenab et al. Eur. J. Lipid Sci. Technol. 108 (2006) 488492

and an FID. Oven temperature started at 190 7C and

increased to the final temperature of 220 7C with a ramp
of 1 7C/min and was held for 10 min. Injector and detector
temperatures were set at 220 and 250 7C, respectively.
Nitrogen gas was used as the carrier. Iodine values and
unsaponifiable matter were measured using methods
Cd 1e-85 and Ca 6a-40 from AOCS [15], respectively.
Chlorophyll concentration was determined using method
Cc 13d-55 [15] using a UV-Visible spectrophotometer
from Varian Corporation, model Cary 50 Scan, based on
spectrophotometric absorption measurements at 630,
670 and 710 nm. Triplicate samples were used for the
analysis of the above parameters.

Fig. 2. Extraction of canola oil with supercritical CO2 at

34.0 MPa and 40.0 7C.
2.5 Statistical analyses

The General Linear Model (GLM) procedure of the Statis-

In agreement with the results of this study, Bulley et al. [6]
tical Analysis System (SAS) release 8.2 (SAS Institute,
observed that in the SCCO2 extraction of crushed canola
Cary, NC, USA) was used for data analysis. Means and
seeds at 35 MPa and 40 7C, the oil solubility in SCCO2 was
least significant differences (LSD, 95% confidence level)
7.2 mg oil/g CO2 at a CO2 flow rate of 1.84 g/min. Also,
were determined to evaluate the differences between the
Fattori et al. [9] reported that in the SCCO2 extraction of
yields, fatty acid compositions, iodine values, unsaponi-
flaked and cooked canola seeds, the oil concentration of
fiable matter and chlorophyll concentrations of the oils
SCCO2 was 11.0 mg oil/g CO2 at a CO2 flow rate of 0.7 g/
extracted by SCCO2 and those by AW406 solvent.
min at 36 MPa and 40 7C, which is relatively higher than
the results of this study. The difference in the operating
pressures from 34.0 MPa in this study to 36 MPa by Fattori
3 Results and discussion et al. [9] cannot justify such an increase in the solubility.
Also, the results reported by Fattori et al. [9] are not con-
3.1 Oil solubility sistent with those of Bulley et al. [6]. Furthermore, Bulley et
al. [6] also reported a slightly higher solubility (8.7 mg oil/
Fig. 2 presents the extraction curves of crushed and
g CO2) when a lower CO2 flow rate (0.64 g/min) was
cooked canola seeds at 34.0 MPa and 40 7C with a CO2 flow
applied, which was mainly due to the better recovery of
rate of 1.6 6 0.2 L/min (after expanding under atmospheric
the extracted oil at the lower CO2 flow rate in the collection
conditions). The slope of the extraction curves were used to
vessel. According to Rezaei and Temelli [16], within
obtain the oil solubility, which is proportional to the mass
experimental errors there are no differences in the oil
transfer rate, under the conditions of this study and was
solubilities with an increase in the flow rate from 0.5 to
measured at 7.1 mg oil/g CO2. According to Rezaei and
4.0 L/min (under atmospheric conditions). Therefore, the
Temelli [16], throughout a batch operation, extraction kinet-
results reported by Fattori et al. [9] cannot be related to the
ics includes three consecutive phases: (1) The linear phase,
lower CO2 flow rate in their study (0.7 g/min, 0.36 L/min
the slope of which can be applied to obtain the solubility of
under atmospheric conditions). Although the flow rate
the extracted lipids at the pressure and temperature of the
should not have such an effect on the oil solubility, in
study. In this phase, oil is freely available on the surfaces of
practice small differences can occur due to a difference in
the solid particles where it can be easily removed. (2) When
matrix structure as well as the possible oil losses in the
the free oil supply is minimized and saturation of the super-
system at higher CO2 flow rates. No sample was collected
critical solvent can no longer be maintained, oil diffusion
when a second trap was used for the collection of possible
from the internal structure of the solid matrix starts to have a
oil carried away from the collection vessel in this study.
greater contribution in the extraction kinetics, and as a result,
the extraction curve enters into a transition phase, where oil
solubility slowly loses its role in the overall extraction kinet- 3.2 Extraction yield
ics. (3) On the last portion of the extraction curve, a plateau is
observed, which indicates that the extraction kinetics is Under the conditions of this study, the apparent yield from
exclusively controlled by the diffusion rate of the lipids SFE was 20.2 6 0.4% (wt/wt, dry-based), which was
through the solid matrix [16]. lower than that of the extraction with organic solvent

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Eur. J. Lipid Sci. Technol. 108 (2006) 488492 Quality comparison of crude canola oils 491

(34.2 6 1.6%). Due to the incomplete SCCO2 extraction Tab. 2. Fatty acid compositions (%) of oils extracted from
process in this study, extraction yields for the SCCO2- crushed and cooked canola seeds using SCCO2 and
extracted runs were lower (runs above 6 h long were diffi- AW406, in the order of their appearance on the chroma-
cult to operate with the system applied in this study). togram.
Canola meal removed from the extraction cell after the
extraction was over indicated that there was oil left on the Fatty acid SCCO2- Solvent-
surfaces of the samples. Also, considering economical extracted oil* extracted
aspects of oil extraction, a longer operation time may not oil*
be justified. Despite those issues, SFE can offer extra
16:0 3.8 6 0.1a 3.7 6 0.1a
advantages over the organic solvents. SFE can provide 18:0 2.0 6 0.1a 2.1 6 0.1a
more selective extraction and also the extracted oil car- 18:1 67.0 6 0.1a 66.9 6 0.3a
ries higher premiums compared to the extraction with 18:2 17.3 6 0.2b 16.9 6 0.1a
organic solvents. Oil extracted by organic solvents con- 18:3 8.0 6 0.1a 7.9 6 0.1a
tains numerous undesirable components that require 20:0 0.6 6 0.1a 0.7 6 0.1a
extra processing steps for their removal. Refining losses 20:1 1.3 6 0.1a 1.3 6 0.1a
with organic solvents have also been reported by many 22:0 Trace 0.2 6 0.1
22:1 Trace 0.4 6 0.1
studies [7, 11, 12]. Furthermore, environmental issues and
SFA 6.5 6 0.1 6.7 6 0.3
health concerns related to the use of many organic sol- MUFA 68.3 6 0.1 68.5 6 0.1
vents for processing a food commodity are among the PUFA 25.3 6 0.1 24.8 6 0.2
drawbacks those solvents are suffering from. Therefore, Iodine value 109.4 6 0.2a 108.7 6 0.5a
considering these issues, it is worthwhile to invest in the
SFE of edible oils and avoid the problems associated with 16:0: palmitic acid; 18:0: stearic acid; 18:1: oleic acid;
the use of organic solvents such as hexane and AW406. 18:2: linoleic acid; 18:3: linolenic acid; 20:0: eicosanoic
acid; 20:1: eicosenoic acid; 22:0: behenic acid; 22:1:
erucic acid; SFA: saturated fatty acids; MUFA: mono-
unsaturated fatty acids; PUFA: polyunsaturated fatty
3.3 Chemical characterization of extracted oils
acids.
*
Mean 6 standard deviation from three extractions under
Fatty acid compositions of the oils extracted by SCCO2
the same operating conditions. In each row, means with
and AW406 are shown in Tab. 2. According to the results,
the same letter are not significantly different (p .0.05).
the concentrations of erucic and behenic acids in the oils
extracted by SCCO2 were minimal compared to those
extracted using AW406. However, the linoleic acid con-
centration of the oil extracted with SCCO2 was somewhat justified, which also verifies the higher selectivity of SFE
higher. The mass transfer rates of fatty acids in SCCO2 are compared to extraction with organic solvents. Iodine
controlled by their chain length and the saturation level of values of the oils extracted by SCCO2 and AW406 are
their chemical bonds. As a consequence of that, at the also shown in Tab. 2. No significant differences (p .0.05)
beginning of the extraction with SCCO2, the triacylglycer- were observed between the iodine values of oils extract-
ols high in PUFA were extracted at higher rates than those ed by SCCO2 and AW406. Since the iodine values repor-
containing long-chain fatty acids such as erucic and ted in this study were directly calculated from the fatty
behenic acids. As reported by Fattori et al. [11], both high- acid profiles of the oils extracted by the two methods,
and low-molecular-weight triacylglycerols are equally which were very similar to each other, such similarity in
soluble in hexane and therefore no selective extractions the iodine values of the oils can also be justified.
can be performed when using organic solvents for oil
extraction. Snyder et al. [17] observed that the last 10 The concentrations of chlorophyll and unsaponifiable
15% of soybean oil extracted by SCCO2 had lower levels matter from the two extracted oils are shown in Tab. 3.
of linoleic, linolenic and palmitic acids and higher levels of The chlorophyll concentration in the oil extracted with
oleic and stearic acids compared to the rest of the frac- SCCO2 was significantly lower than that obtained by
tions. Similarly, Fattori et al. [11] reported that the last AW406 solvent. This is due to the lower solubility and/or
20% of canola oil obtained during SCCO2 extraction mass transfer rate of polar components such as pig-
showed higher concentrations of behenic and lignoceric ments, including chlorophyll, in SCCO2. However, such
acids. Because of the incomplete extraction process in pigments can be extracted at higher CO2 volumes, when
this study, the last portions of the extraction process did other oil components have already been taken away [12].
not exist in the oil extracted with SFE. Therefore, the trace Therefore, the incomplete SCCO2 extraction, as occurred
amounts of behenic and erucic acids in this study can be in this study, can result in an oil low in polar components

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492 E. Jenab et al.
such as the pigments. Consistent with this study, the
studies of Friedrich et al. [7] and List et al. [18] on soybean
Tab. 3. Comparison of SCCO2- and AW406-extracted
crude canola oils. The concentrations are calculated
based on the total oils extracted.
Analysis AW406* SCCO2*
Chlorophyll [ppm] 8.2 6 0.1a 6.0 6 0.7b
Unsaponifiable matter [%] 1.1 6 0.1a 1.1 6 0.1a
* [4] F. Temelli, N. T. Dunford: The effect of processing parame-
Mean 6 standard deviation from three extractions under
the same operating conditions. In each row, means with
the same letter are not significantly different (p .0.05).
and cottonseed oils, respectively, showed that oils
extracted with SCCO2 were lower in pigments than oils
extracted by organic solvents. As observed in this study,
Friedrich and Pryde [8] also indicated that cottonseed,
wheat germ and bran oils extracted by SCCO2 were
lighter in color, which was due to the lower concentration
of pigments in such extracts. The seed flakes packed in a
long cylindrical extractor act like a stationary phase in the
chromatographic columns, and as a result, less polar
compounds such as triacylglycerols can move faster
through the seed phase than more polar compounds
such as pigments and phospholipids [2, 8, 12].
No significant differences (p .0.05) were observed be-
tween the amounts of unsaponifiable matter of the
extracted oils obtained by SCCO2 and AW406 (Tab. 3).
Consistent with this study, Friedrich et al. [7] reported that
there were no significant differences in the total unsapo-
nifiable matter between SCCO2- and hexane-extracted
soybean oils. However, Friedrich and Pryde [8] reported
that the amounts of unsaponifiable matter of SCCO2-
extracted oils from wheat germ and bran were lower than
those obtained using organic solvents. Friedrich et al. [7]
and Przybylski et al. [12] reported that unsaponifiable
matter was not extracted at a constant rate when using
SCCO2 for the extraction.
Acknowledgments
This work was funded by a grant provided by the Office of
the Vice-President for Research in the Campus of Agri-
culture and Natural Resources at the University of Tehran.
2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Eur. J. Lipid Sci. Technol. 108 (2006) 488492
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[Received: February 2, 2006; accepted: March 31, 2006]
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