122 DOI 10.1002/ejlt.200300891 Eur. J. Lipid Sci. Technol.

106 (2004) 122–125

Pavan K. Mamidipally First approach on rice bran oil extraction using

Sean X. Liu
limonene
Department of Food Science,
Rutgers, Edible oil extraction with petroleum derivatives as solvents has caused safety, health,
The State University and environmental concerns everywhere. Thus, finding a safe alternative solvent will
of New Jersey, New Brunswick, have a strong and positive impact on environments and general health of the world
NJ, USA population, considering the scale of oil extraction operations worldwide. The extrac-
tion of oil from rice bran by d-limonene and hexane (for comparison) has been carried
out at their respective boiling points at various solvent-to-meal ratios and for various
extraction times. The preliminary data suggested that the optimum solvent-to-meal
ratio and extraction time required for d-limonene extraction of rice bran oil to be 5:1
and 1 h respectively. The initial quality characteristics (free fatty acid content, oil color,
phospholipid content) of crude oil extracted under these optimum conditions were an-
alyzed using various analytical methods based on the standard methods of AOCS and
were found to be comparable to the oil extracted with hexane. The initial positive result
has paved the way for further studies on issues related to meal qualities as well as to a
scale-up of the method in the near future.

Keywords: Alternative solvent, d-limonene, edible oil extraction, hexane, rice bran.

1 Introduction hexane emissions into the environment every year. This

Commercial-grade hexane, a paraffinic petroleum frac-
tion, has been the solvent of choice worldwide for extrac-
tion of oil from oilseeds for both practical and economical
reasons. It has a fairly narrow boiling point range of ap-
proximately 63–697C and is an excellent oil solvent in
terms of oil solubility and ease of recovery. However,
such desirable properties of hexane are also directly
responsible for serious problems such as hexane escape,
fire, explosion, and air pollution. Residual hexane in the
defatted meals has been found to be toxic to piglets and
the meals are banned as animal feeds in a number of mar-
Short Communication
would easily surpass the emission limits and thus most
of these oilseed companies end up paying huge fines
and are required to get a Federal Operating Permit.
All these concerns have stimulated the interest in the re-
search on alternatives to hexane as an extraction solvent.
In the past the research on alternative solvents had
focused on ethanol, isopropyl alcohol, halogenated
hydrocarbons, hydrocarbons, sulfur containing solvents,
oxygenated hydrocarbon solvents and even water [1–4].
A comprehensive review on the alternative solvents
explored in the past can be found in the literature [5].

kets. Hexane, when inhaled by humans, is known to dis-
solve in neural lipids thus affecting the nervous system. n-
Hexane, the main component of commercial hexane, is
listed as the No. 1 of 189 hazardous air pollutants (HAPs)
by the US Environmental Protection Agency, with the
edible oil processing industry being listed as a source
category for hexane emissions. The industry is consid-
ered a major source in case it emits more than 10 t per
year of HAPs, thus requiring a US Federal Operating Per-
mit [4]. Even when the moderate loss of 0.15% per ton of
feed is considered, a plant with an average daily proces-
sing capacity of about 75 t per day would result in 42 t of
Correspondence: Sean X. Liu, Department of Food Science,
Rutgers, The State University of New Jersey, 65 Dudley Road,
New Brunswick, NJ 08901, USA. Phone: +1-732-932-9611-240,
Fax: +1-732-932-6776; e-mail: liu@aesop.rutgers.edu
The alternate solvent investigated in this study is limo-
nene (with d-limonene as the main component) that is
the major component of the oil extracted from citrus rind.
d-Limonene has the chemical name (4R)-(1)-4-isoprope-
nyl 1-methylcyclohexane and its chemical structure is
shown in Fig. 1. At 25 7C, citrus (1)-limonene can rotate
the plane of polarized light at least 967 to the right (the
“d” comes from the word, “dextro-rotatory”, an older des-
ignation for the isomer’s ability to rotate a plane of polar-
ized light to the right (1)). This increases with the purity of
d-limonene. d-Limonene is considered as GRAS (Gener-
ally Recognized as Safe) material by the U.S. Food and
Drug Administration. d-Limonene has been used as
cleaning/degreasing product in both industrial use and in
household applications. It is capable of cleaning organic
dirt loads ranging from light cutting oils and lubricants to
the heaviest greases. The relevant properties of d-limo-

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Eur. J. Lipid Sci. Technol. 106 (2004) 122–125 Edible oil extraction with d-limonene 123

edible oil and nutraceutical industries. In this short com-

munication paper, the preliminary study on extraction
characteristics of d-limonene is described and the results
are compared to the extraction characteristics resulting
from hexane extraction.

Fig. 1. Chemical structure of d-limonene [7].

2 Materials and methods

nene as compared to hexane as a solvent are listed in 2.1 Materials

Tab. 1 [3, 7]. Hexane comes from the same feedstock as
Extrusion-stabilized rice bran in the form of collets (pel-
gasoline. Hence its price is determined by the supply and
lets), donated by Riceland Foods USA (Stuttgart, Arkan-
demand for gasoline. Limonene is not as plentiful as hex-
sas, USA), was used as a model oilseed for extraction
ane and the price of limonene is therefore higher than that
studies. Food-grade d-limonene (96% purity) was pur-
of hexane. However, taking into account the costs asso-
chased from Florida Chemical Company (Winter Haven,
ciated with environmental compliances and insurances,
Florida, USA). Hexane and all the other chemicals were
the limonene extraction process is competitive. Further-
purchased from Fisher Scientific (Atlanta, USA). Initial
more, many health-promoting lipophilic nutraceuticals
moisture content of rice bran was 5.97% and oil content
(e.g. carotenoids, lycopene) have been extracted with
was determined to be 28.42% according to the AOCS
hexane, which, though a far cry from using such toxic sol-
method Ac 3-44 [8]. Rice bran was stored in the freezer
vent as benzene, is still unsettling since these health
until used for the extraction to prevent any lipid oxidation.
products may contain residual hexane. It is highly likely
Prior to the extraction, rice bran was ground using a mor-
that consumers would prefer those nutraceuticals pro-
tar-pestle and the ground bran was sieved using USA
cessed with non-toxic and natural solvents. Ultimately,
standard 8 and 70-mesh screens to remove the oversize
the feasibility of limonene as a solvent for food and nutra-
and fines.
ceutical industries is determined by many factors, includ-
ing economical and environmental factors. The primary
objective of this stage of the research was to determine if 2.2 Extraction of rice bran oil with solvents
d-limonene could be used as an alternate solvent in
A known quantity of ground rice bran (25 g) was extracted
with hexane or d-limonene at their respective boiling
Tab. 1. The relevant properties of d-limonene as a sol-
vent compared to hexane [3, 7]. points. The extraction was carried out at various solvent-
to-bran ratios (wt/wt) of 2:1, 3:1 and 5:1 and times of
Property Hexane d-Limonene extraction of 0.5 h, 1 h, 2 h and 3 h. After extraction, the
miscella from each flask was separated from the defatted
Molecular weight 86.17 136.23 rice bran by vacuum filtration. Vacuum evaporation, using
[kg/kg mol]
rotary vacuum evaporator, of solvent from the miscella
Specific gravity [298 K] 0.65 0.84 yielded crude rice bran oil. But in the case of d-limo-
Viscosity [Pa.s] [298 K] 0.00032 0.00092 nene-extracted oil, the miscella was subjected to further
Boiling point [K] 341.886 435.930 evaporation in a vacuum oven at 82 7C for 2 h and maxi-
Latent heat of 334.302 353.129 mum vacuum to remove the remaining d-limonene. The
vaporization [kJ/kg] conditions for evaporation in the vacuum oven were
Specific heat [kJ/kg.K] 2.230 1.832 achieved after several trials using different temperature
and vacuum combinations and the use of gas chromatog-
Solubility in water 0.05 0.01
[kg/m3] [298 K] raphy to make sure that there was no d-limonene in the
residual oil after evaporation. The crude rice bran oil yield
Dielectric constant 1.89 2.37
[293 K] was then measured gravimetrically.

Flash point [K] 250.150 321.150 Having tentatively determined the optimal conditions for
Surface tension 0.018 0.027 extraction, the quality of crude oil was analyzed for both
[N/m] [298 K] 5:1/1 h d-limonene-extracted oil and the 3:1/1 h hexane-
Renewable No Yes extracted oil. Phosphorus content of the weighed amount
Toxic Yes No of crude oil was determined according to the AOCS meth-
od Ca 12-55 [8]. The acceptable conversion factor of 31.7

 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.ejlst.de

124 P. K. Mamidipally and S. L. Liu Eur. J. Lipid Sci. Technol. 106 (2004) 122–125

[9] was then used to convert the phosphorus values to rice bran oil respectively. As can be observed from Tab. 2,
phospholipids. The total free fatty acid content in the d-limonene extracted a significantly higher amount of oil
crude oil was determined according to the AOCS method than hexane under any given condition. Moreover it can
Ca 5a-40 [8] and the values are expressed as percent also be observed that with further increase in the time of
oleic acid. The color of the crude oil was measured in hex- extraction after 1 h for solvent-to-bran ratio of 5:1, the
ane by the spectrophotometric index method [10]. The increase in the oil yield in case of d-limonene was statisti-
absorbance of the crude oil (5% crude oil in hexane) was cally insignificant while 2:1/2 h and 3:1/1 h seem to be the
measured against hexane as reference at the wavelength optimum conditions for hexane extraction. Bhagya et al.
of 430 nm in a 1-cm cuvet. [12] have reported that for soybean extraction carried out
for solvent-to-flakes ratio of 1.5:1, further increase in
extraction time after 1 h resulted in significant increase in
2.3 Statistical analysis the phospholipid content of the extracted oil leading to
heavy refining losses. It can be concluded that 5:1/1 h for
The data were analyzed using the SAS general linear
d-limonene and 3:1/1 h for hexane were the optimum
models (GLM) procedure (SAS Institute, Cary, North Car-
conditions for optimal yield of crude oil for the respective
olina, USA). An alpha value of 0.05 was used to test if
solvents.
there is any significant difference between the data for
the respective solvent. Tab. 3 summarizes the quality characteristics of the crude
oil extracted for 5:1 ratio/1 h d-limonene and 3:1 ratio/1 h
hexane-extracted crude oils. The percentage of total free
3 Results and discussion fatty acids and phospholipid content of d-limonene
extracted oil were comparable to that of hexane-
The crude rice bran oil yields were expressed as wt-% of
extracted oil. The values are slightly higher in the case of
the dry full fat rice bran used and are tabulated in Tab. 2.
d-limonene-extracted oil compared to that of hexane.
For hexane extraction, carried out for 1 h, an increase in
This may be due to the slightly polar nature of d-limonene
the solvent-to-bran ratio (wt/wt) from 2:1 to 3:1 yielded
when compared to hexane. The color of the d-limonene-
15.2% more crude rice bran oil while the increase was
extracted oil (as measured by absorbance value) was
only 1% from 3:1 to 5:1. Hu et al. [11] reported a 10.8%
found to be slightly darker compared to the hexane
increase when the solvent-to-bran ratio was raised from
extracted oil as indicated by higher absorbance values
2:1 to 3:1 with an extraction carried out for 0.5 h at 60 7C.
even though visually there was not much difference be-
On the other hand for d-limonene extraction carried out
tween the two. The reason of d-limonene-extracted oil
for 1 h, an increase in solvent-to-bran ratio from 2:1 to
being slightly darker compared to hexane-extracted oil
3:1 and 3:1 to 5:1 produced 9.8% and 14.9% more crude
might be due to the higher temperature used during
extraction and evaporation when d-limonene is used as
Tab. 2. Rice bran crude oil yield (wt/wt) after solvent
extraction. the solvent. This might be overcome by using vacuum to
lower the temperature of evaporation and membrane
Solvent-to- Time Oil extraction yield{ [%] separations for solvent recovery. The oxidation stability
meal ratio [h] of d-limonene during the extraction-recovery cycle is
d-Limonene Hexane
being tested; the preliminary data indicate that the oxi-
2:1 0.5 15.8 6 0.2a 14.4 6 1.1c dized products of d-limonene after one cycle contain
2:1 1 18.3 6 0.1d 15.8 6 1.5b, c
2:1 2 18.9 6 0.5c, d 17.0 6 0.4b
2:1 3 19.2 6 0.3c, d 17.3 6 0.7b Tab. 3. Quality characteristics of d-limonene and hex-
3:1 0.5 19.2 6 0.2c, d 15.7 6 0.1b, c ane-extracted crude oil.
3:1 1 20.1 6 0.7c, d 18.2 6 0.6b
Quality characteristics d-Limonene Hexane
3:1 2 21.1 6 0.1c 18.4 6 0.9b
5:1, 1 h 3:1, 1 h
3:1 3 21.1 6 0.1c 18.9 6 0.4b
5:1 0.5 20.7 6 0.1c 17.3 6 0.5b Free fatty acid{ 2.51 6 0.29 2.05 6 0.12
5:1 1 23.1 6 0.9b 18.2 6 0.4b (as oleic acid) [wt-%]
5:1 2 24.5 6 2.5b 18.4 6 1.5b
5:1 3 24.6 6 1.6b 18.6 6 0.1b Phospholipids{ [%] 1.50 6 0.25 1.37 6 0.07
Color (430 nm, 5% oil in 0.31 6 0.02 0.08 6 0.01
{
The values are means of duplicate determinations hexane), absorbance{
6 standard deviation. Significantly different values
{
(P ,0.05) in the same column are indicated by different The values are the means of two independent determi-
letters. nations 6 standard deviation.

 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.ejlst.de

Eur. J. Lipid Sci. Technol. 106 (2004) 122–125
about 2443 ppm of cis/trans-limonene-1, 1336 ppm of
carveol, 539 ppm carvone, from originally 1835 ppm,
657 ppm, and 429 ppm, respectively, without using either
vacuum for extraction or adding antioxidants such as the
mixture of butylated hydroxyanisole (BHA) and butylated
hydroxytoluene (BHT), to the extractor.
To summarize, the optimal conditions for crude rice bran
oil extraction with d-limonene are a 5:1 solvent-to-bran
ratio (wt/wt) and a 1-h extraction time, whereas the quality
of crude oil extracted under these conditions was found
to be comparable to that of hexane. The extent of oxida-
tion of d-limonene used for extraction is being studied at
this moment. Our current limited data indicate that the
oxidation of limonene during an extraction-recovery cycle
was insignificant as the oxidized limonene represents less
than 1 wt-% of d-limonene. Hence it may be tentatively
concluded that d-limonene is suitable for reuse as a sol-
vent. The major potential trade-off of using d-limonene
instead of hexane in edible oil/nutraceutical extraction is
the higher energy consumption related to solvent recov-
ery due to the higher boiling point and to higher specific
gravity when compared to hexane. The price of food
grade limonene is higher than that of hexane (about 2:1
depending on the price of crude petroleum in the world
market), however, if the factors related to regulatory com-
pliance costs, costs of workers’ health, as well as those of
the environment are included in the final analysis of pro-
cess economics, the outcome is still to be determined. It
is therefore necessary to carry out a pilot scale testing in
the next 6 months to provide data for economical analysis
of d-limonene as a solvent for lipo-soluble material recov-
ery. Further studies on defatted meal quality, scale-up,
peroxide value of the oil, and membrane separation of oil
from the miscella will be carried out. It is also to be deter-
mined if d-limonene is suitable to extract more vitamin E
and oryzanols (potential antioxidants) because of its
slightly polar nature compared to the highly nonpolar hex-
ane. The research has additionally broader implications for
the extraction of other lipo-soluble neutraceuticals from
agricultural and food by-products. This includes the
extraction of lycopene, a lipo-soluble phytochemical with
a promising health-promoting property. Hexane is often
used in the laboratory for the extraction of lycopene, how-
ever due to health concerns with the residual hexane in
lycopene and safety associated with the production, a
more environmentally friendly solvent is highly desirable
[13]. Hence d-limonene might be an attractive alternative.
 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Edible oil extraction with d-limonene 125
Acknowledgements
The authors would like to thank New Jersey Agricultural
Experiment Station at Rutgers University for funding this
work and Riceland Foods USA for providing rice bran
samples.
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[Received: August 4, 2003; accepted: November 17, 2003]
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