───Food Technology ───

Physicochemical composition and FTIR characterization

of castor seed oil
Tarique Panhwar1, Sarfaraz Ahmed Mahesar1, Aftab Ahmed Kandhro2,
Syed Tufial Hussain Sheerazi1, Abdul Hameed Kori1,
Zahid Hussain Laghari1, Jamil-ur-Rehman Memon2

1 – National Centre of Excellence in Analytical Chemistry, University of Sindh, Pakistan

2 – Dr. M. A. Kazi Institute of Chemistry, University of Sindh, Pakistan

Abstract
Keywords:
Introduction. This paper provides the complete profile
Oil of physicochemical properties and fatty acid composition
Castor of local castor varieties from Sindh, Pakistan. Initially,
Seed physical characteristics of seeds were studied followed by
FTIR detailed examination of oil.
Materials and methods. Oil was extracted through
Soxhlet extraction method followed by physico–chemical
examination of necessary parameters (iodine value,
peroxide value, saponification value, viscosity and
Article history: moisture). For qualitative analysis of castor oil GCMS and
FTIR instruments were used.
Received 14.02.2019
Received in revised form Results and discussions. Oil content was observed in
25.08.2019 the range of 44–48%. Moisture and ash content of castor
Accepted 28.11.2019 seeds were found to be 4.22–5.16% and 5.66–6.49%,
respectively. Compositional analysis by GCMS has shown
ricinoleic acid (88.5–93.1%) as a prominent fatty acid in all
local varieties. Other fatty acids present were palmitic (0.4–
Corresponding author:
0.8%), stearic (0.8–1.0%), linoleic (2.8–3.3%), and
Tarique Panhwar ecosenoic (0.2–1.5%). Free fatty acid value of all castor oil
E-mail: varieties was found to be in the range of 0.16–0.53%. Other
panhwertarique@ quality parameters such as IV, PV and SV of different
gmail.com castor oil varieties were determined in the range of 79.16–
90.03 gI2/100g, 1.62–1.89 meq/Kg, and 188.12–204.76
mgKOH/g, respectively. The FTIR spectra of castor seed
oil varieties were nearly similar. On careful examination of
the intensity values of each functional group present in the
oil showed some variations in castor seed varieties.
Conclusion. Indigenous local varieties of castor with
sufficient oil content (> 40%) were explored and examined
for some important parameters. FTIR band intensities of
some functional groups highly correlated with important
DOI: 10.24263/2304- parameters of castor oil.
974X-2019-8-4-9

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Introduction

Castor is one of the most promising non-edible oil seed crop with great industrial
importance, grown all over the world in warm temperate, tropical and subtropical regions [1].
Castor belongs to Eurphorbiaceae family and botanically known as Ricinus Communis L. [2].
The purpose of cultivation of castor plant is because of its seeds that have high oil content.
The oil is pale yellow in color, viscous in nature with a lot of applications in industrial and
medicinal fields [3].
Since the fatty acid composition consists of about 90% ricinoleic acid and other fatty
acids include oleic acid 2.8%, palmitic acid 0.7%, stearic acid 0.9%, linoleic acid 4.4%, and
linolenic acid 0.2% [4]. The unique properties of castor oil among other vegetable oils are
due to the ricinoleic acid (a unique fatty acid C18H34O3) known as cis-12-hydroxyoctadeca-
9-enoic acid. Castor oil has relatively high viscosity, specific gravity and solubility in
alcohols due to high hydroxyl value in the nature [5].
Castor oil is widely used as a raw ingredient in the manufacture of pharmaceuticals,
cosmetics, coatings, soap, nylon, lubricants, dyes, inks, and waxes [6]. More over castor oil
is used in various industrial fields as a potential by product [7]. Castor oil has limited food
value and do not compete with food crops [3]. It is mainly cultivated for trading purpose in
more than 30 countries of the world in the area of 1.525 million hectares, which produce 1.58
million tons of seed. Major castor growing countries are India, Brazil, China, Thailand,
Russia, Ethiopia and Philippines [8]. India has major contribution of 65% of castor oil all
over the world, thus fulfils 80 to 90% of world’s requirement of ricinus oil [1].
Pakistan is agricultural country with 70% population belongs to an agriculture sector
[9]. Pakistan has total area of 79.6 million hectares, 70% of which is arid to semi-arid. The
total unused land is about 28 million hectares, due to shortage of water, severe heat and saline
nature of soil, the large area of land is unproductive [9, 10, 11] and therefore castor plant has
potential to grow on marginal lands as compared to other non-edible oil yielding plants [12].
A sufficient research work has been reported on physicochemical characteristics of
castor oil [2,13] and its fatty acid composition [5]. However, a limited work is done on
searching new varieties of castor seed for complete profiling. The current study presents some
more aspects to study new castor varieties, which include complete profiling of castor seed
in the light of environmental factors such as light, altitude, latitude and soil type.
Furthermore, FTIR study was also carried out to characterize castor oil and correlated with
the physiochemical properties.

Materials and methods

Reagents and sample collection

All the chemicals and reagents used in the present work were purchased from E-Merck
(Darmstadt, Germany). Castor beans were collected from fifteen different locations of Sindh
province. Some harvesting sites had different environmental conditions and soil types. Five
locations were selected within the same environmental conditions. The plants at different
sites showed variability in their height. Branch distribution in castor plant was ramified. Sites
1-5 belong to middle region of Sindh province, where luminosity exposure was observed.
The region has low humidity with temperature of 40–50 °C throughout the growing season
and type of soil was non-sodic, non-saline and alluvium in nature having more ratios of silt
and clay with moderate level of water observed at sites. Sites 6–10 belong to downstream of

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Sindh province with the temperature of 35 °C with humidity of about 60%. Soil was rocky,
slightly saline with little water present in the area. Sites 11–15 also come in downstream with
the temperature of 35 °C. Soil was fertile, non-sodic, non-saline. The collected castor seeds
were broken off and packed in clean sampling bags until analysis.

Classification
Castor seeds were classified on the basis of color, plant characteristics, and seed weight.
On the basis of classification, the collected castor seeds generated three new local varieties.
These newer local varieties were coded as S-1, S-2 and S-3. All castor varieties were dried
and cleaned in order to remove some foreign materials and stored at room temperature in the
absence of light.

Oil extraction
Crude oil from castor beans was extracted by Soxhlet extraction method using n-hexane
as a solvent with slight modification as reported by Akpan et al., 2006 [13]. Finely ground
10g of sample were put into thimble and carefully placed inside the Soxhlet extractor, then
300 mL of n-hexane was added to round bottom flask. The process continued for about 6 h
at 65 °C. After complete extraction hexane was evaporated using rotary evaporator (Buchi,
Switzerland). For total oil content, weight of the extracted oil was measured using electronic
balance. The oil was kept at low temperature for further physicochemical analysis.

Physicochemical analysis of seed and oil

Seed characteristics. Some physical characteristics of castor seeds of different varieties
were analyzed such as seed color, weight, thickness, length and weight of 100-seeds. Specific
gravity and refractive index were measured according to reported method [13].Viscosity of
oil samples was measured using a viscometer (VM 3000 stabinger Anton Park, Austria) at
40 °C using 5 mL of sample. Ash content was determined by AOCS (method Ba 5a-49) [14].
Moisture content measurement was done by drying up to constant weight at 105 °C [15].
Chemical analysis. For the chemical characteristics official methods of AOCS were
used for the determination of iodine value (IV method- Cd 1-25), peroxide value (PV method-
Cd 8-53), saponification value (SV method-Cd 3-25) and free fatty acids (FFA method- Aa
6-38)[14].
FTIR analysis of castor oil. For the qualitative analysis of castor oil, infrared (IR)
spectra were recorded using FTIR spectrometer (Thermo Nicolet Avatar 330) with a DTGS
detector (Thermo Nicolet Analytical Instruments, Madison, WI). The removable (ZnSe
crystal) single bounce-attenuated total reflection (ATR) accessory was used for oil analysis
with the conditions as follows: resolution 4 cm -1, scans 32, IR range 4000–650 cm-1. The
background spectrum was recorded before the sample analysis. After recording the spectra
ZnSe crystal was carefully cleaned with hexane to remove any residue of the previous
sample.
Determination of fatty acid composition. Fatty acid methyl esters (FAMEs) were
prepared according to official IUPAC method No: 2.30 [16]. A complete profile of fatty acid
composition including major and minor fatty acids was studied using GC-MS (Agilent 5975
GC-MSD) with chemstation 6890 Scale mode software. FAMEs were analyzed using Agilent
autosampler 7683-B injector (Agilent Technologies, Little Fall, NY, USA), a HP-5MS (5%
phenyl methylsiloxane) capillary column with dimension of 30 m, i.d 0.25 mm. The initial
temperature was set as 150 °C and maintained for 2 min then raised to 220 °C at a ramp of 3

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°C
/min and finally held for 15 minutes. Carrier gas used was helium at a flow rate of 1.5
mL/min. About 2-µL of FAMEs were injected into the column using split mode injection
system. The mass spectrometer was operated using electron impact mode at 70eV in the scan
range of 50-550 m/z. GC-MS chromatograms were compared with two libraries (NIST and
Wily) which provided much information about major and minor fatty acids in castor oil.

Statistical analysis
Two samples of each variety (1 kg each) were collected and every sample was analyzed
thrice and the obtained results were reported as mean±standard deviation, by using
Microsoft® excel 2003 software.

Results and discussion

Table 1A shows the geographical locations of three different castor seed harvesting
sites. S-1 variety was collected from sites 1–5, S-2 variety from sites 6-10 and S-3 variety
from sites 11–15. Table 1B represents physical characteristics of three different castor seed
varieties. Some variations among different varieties were found; this might be due to different
environmental conditions and soil types [17]. Moisture and ash content of castor seeds
showed little variability as shown in Table 1B. Moisture and ash content of castor seeds were
found to be 4.22–5.16% and 5.66–6.49%, respectively.
Table 2 shows the proximate composition along with some quality attributes of different
castor varieties. A significant variability was found in the oil content of studied castor seed
varieties that ranged from 44–48%. The obtained values showed less oil content than
indigenous variety reported earlier [18]. Observed variations in the seed oil content may be
due to castor seed genotypes, harvesting time, environmental and cultural aspects.
Refractive index, specific gravity and viscosity in castor varieties were found in the
range of 1.4321–1.4532, 0.9531–0.9576 g/cm-3 and 663–713 mPas.s, respectively. The high
viscosity of castor oil is due to the presence of long chain carbon atoms which mainly consist
of ricinoleic acid; a mono-saturated, 18-carbon fatty acid having hydroxyl group at 12th
carbon, a very uncommon property for a biological fatty acids which makes it unique oil from
other vegetable oils.
The chemical parameters such as IV, PV and SV of different varieties were determined
in the range of 79.16–90.03 gI2/100g, 1.62–1.89 meq/Kg, and 188.12–204.76 mgKOH/g,
respectively. The quality of oil depends on the presence of FFA. Generally these values are
calculated as percentage of oleic acid. In current study, the range of FFA in castor varieties
was determined as 0.16–0.53%. These values are almost comparable to freshly processed
vegetable oils [19].

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Table 1A
Site coordinates where the castor beans were harvested

Site Elevation (m) Latitude Longitude

1 24 26.73° N 67.78° E
2 42 26.41° N 67.37° E
3 43 26.56° N 67.71° E
4 43 26.86° N 67.78° E
5 41 26.42°N 67.86°E
6 38 25.41°N 68.27°E
7 29 25.90°N 68.24°E
8 39 25.35°N 68.26°E
9 39 26.03°N 68.13°E
10 38 25.28°N 68.15°E
11 24 25.38°N 68.36°E
12 25 25.42°N 68.54°E
13 21 25.52°N 69.01°E
14 22 25.45°N 68.72°E
15 26 25.76°N 68.66°E

Table 1B
Physical characteristics of castor seeds

Mean±SD
Seed 100-seed
Seed Qualitative Moisture Ash Length Thickness Width
weight weight
Variety description (%) (%) (mm) (mm) (mm)
(g) (g)
4.22 5.88 11.55 5.47 7.62 0.40 38.86
S-1 Brownish ±0.21 ±0.23 ±0.39 ±0.40 ±0.31 ±0.08 ±0.14
Light 5.16 6.49 12.19 5.20 7.21 0.34 35.18
S-2 ±0.15 ±0.19 ±1.02 ±0.50 ±0.96 ±0.04 ±0.20
brown
Reddish 4.46 5.66 12.55 6.10 7.88 0.48 39.60
S-3 ±0.12 ±0.12 ±1.49 ±0.75 ±0.58 ±0.12 ±0.10
brown

Table 2
Proximate composition and quality attributes of castor varieties

Specific
Oil Viscosity
gravity RI at 28 FFA IV PV SV
Variety content at 28 °C
at 28 °C °C % gI2/100g meq/Kg mgKOH/g
% (mPas.s)
(g/cm-3)
48 0.9531 681 1.4321 0.16 80.34 1.89 197.35
S-1 ±1.21 ±0.02 ±9.11 ±0.05 ±0.006 ±1.55 ±0.05 ±1.81
44 0.9576 713 1.4385 0.27 90.03 1.87 204.76
S-2 ±1.38 ±0.03 ±8.59 ±0.04 ±0.01 ±1.33 ±0.06 ±2.13
46 0.9563± 663 1.4532 0.53 79.16 1.62 188.12
S-3 ±1.22 0.01 ±8.87 ±0.06 ±0.02 ±1.12 ±0.03 ±1.93

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Knowledge of fatty acid profile is much essential for industrial sectors including energy,
cosmetic and soap making industries [20]. Table 3 represents the fatty acid composition of
extracted oil of castor seed varieties. Analysis showed that the most abundant fatty acid in
each variety was ricinoleic acid; found as 88.5%, 90.9% and 93.1% respectively in S-1, S-2
and S-3. Other fatty acids present were palmitic (0.4-0.8%), stearic (0.8–1.0%), linoleic (2.8-
3.3%), and ecosenoic (0.2–1.5%). Three different isomers of oleic acid were also detected in
castor varieties such as oleic n-9 (2.6–4.3%), oleic n-10 (0.4–0.5%), and isomeric oleic n-11
was also found at 0.2%. Ricinoleic acid is the prominent fatty acid found in castor varieties.
The obtained values of this unique fatty acid are even higher than Malaysian (84.2%),
Egyptian (81.9%), and Brazilian (90.2%) castor varieties. Whereas all studied varieties
showed lower recinoleic acid content than Indian castor varieties. Presence of high content
of ricinoleic acid represents good quality of oil.

Table 3
Fatty acid composition of extracted castor seed oil from three varieties of Sindh Pakistan

Variety C16:0 C18:0 C18:1 n9 C18:1 n10 C18:1 n11 C18:2 n9, 12 C20:1 n11 C18:1 n9,12-OH
S-1 0.8±0.02 0.9±0.01 4.3±0.1 0.5±0.02 - 3.3±0.1 1.5±0.05 88.5±3.1
S-2 0.7±0.01 1.0±0.03 3.8±0.09 0.4±0.01 - 3.2±0.1 - 90.9±2.9
S-3 0.4±0.01 0.8±0.02 2.6±0.1 - 0.2±0.01 2.8±0.09 0.2±0.01 93.1±3.3

Table 4 represents the comparative data of fatty acid profile of current varieties with
Malaysian, Indian, Brazilian, Egyptian and indigenous castor variety from this region. The
two fatty acids including isomers of oleic acid (C18:1 n10) and ecosenoic acid (C20:1 n11)
were present in indigenous varieties but both were absent in varieties of other countries. The
total unsaturated fatty acid content was higher than Malaysian, Brazilian, Indian and Egyptian
varieties.

Table 4
Comparison of fatty acid composition of castor varieties with world varieties
Current
Fatty acid (%) Indigineousa Malaysiab Brazilc Indiad Egypte
Study
Palmitic; C16:0 0.4-0.8 0.31 1.3 0.7 - 1.6
Stearic; C18:0 0.8-1.0 0.45 1.2 0.9 1.0 3.4
Oleic; C18:1 n9 2.6-4.3 2.05 5.5 2.8 - 8.3
Oleic; C18:1 n10 0.4-0.5 0.22 - - - -
Oleic; C18:1 n11 0.2 - - - - -
Linoleic; C18:2 n9,12 2.8-3.3 1.84 7.3 4.4 4.3 4.7
Linolenic; C18:3 - - 0.5 0.2 - -
Ricinoleic; C18:1 n9, 88.1-
94.59 84.2 90.2 94.0 81.9
12-OH 93.1
Ecosenoic; C20:1 n11 0.2-1.5 0.53 - - - -
Saturated fatty
1.5-1.7 0.76 2.5 1.6 1.0 5.0
acids (SFA)
Unsaturated fatty 98.1-
99.23 97.5 97.6 98.3 94.9
acids (UFA) 98.9
a b c d
Panhwar et al. (2016) [18], Salimon et al. (2010) [2], Conceicao et al. (2007) [4], Gupta et
al. (1951) [23], eAlgharib and Kotb (2013) [24].

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FTIR Analysis of Castor Seed Oil

The FTIR spectra of castor varieties were nearly similar and no significant difference
served as shown in Figure 1. For the identification of functional groups and bands
corresponding to various stretching and bending vibrations, which clearly indicates the
different characteristic bands of the expected functional groups. The band at 3424 cm -1
represents the O-H stretching vibrations which clearly indicate the presence of hydroxylated
ricinolic acid (prominent peak of castor oil). The band at 3007 cm -1 represents -C-H stretching
vibrations of cis- double bond of unsaturation, whereas the band at 2922 and 2855cm -1 are
characteristics of assymetrical and symmetrical vibrations of aliphatic –CH2 fatty acid
hydrocarbon chain [21,22]. The ester carbonyl (C=O) functional group shows characteristic
stretching band of triglyceride at 1739 cm -1. The bending vibrations of –CH2 scissoring
aliphatic groups observed at 1455 cm -1, while the band at 1366 and 1235 cm-1 are due to
bending vibration of –CH2 groups. The bands at 1235, 1160, 1089 and 1040 cm-1 belongs to
bending vibrations of ester carbonyl group. The band at 971 cm-1 reveal the presence of–
HC=CH-(trans) bending out of plane. The band at 858 cm-1 is for =CH2 wagging vibrations.
However the band at 719 cm-1 is due to overlapping of –CH2 rocking out of plane vibration
of cis-disubstituted olefins, characteristics of long chain fatty acids.

Figure 1. FTR Spectra of three castor seed oil varieties

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Correlation of IR band intensity with physicochemical properties of castor oil

As mentioned earlier that no significant differences was observed in the spectra of castor
seed varieties. On careful examination of the intensity values of each functional group present
in the oil showed some variations in castor seed varieties as shown in Table 5. Therefore,
attempt was made to correlate the intensities of different functional groups with the
physiochemical properties of castor oil. The band at 3421cm -1 showed correlation with the
values of specific gravity as it can be seen from Table 2. The highest specific gravity was
observed in S-2 and lowest in S-1, similarly higher intensity was observed in S-2 and lower
in S-1. FFA showed correlation with the intensity of band at 3007cm-1 as it can be seen from
Table 2, the highest FFA was noted in S-3 and lower in S-1. Usually band at 1709 cm-1 has
been reported for FFA determination. As this band is completely absent in all castor varieties,
therefore band at 3007 cm-1 was selected. The band at 1742 cm-1 was linked with the SV of
castor oil. Higher and lower SV was observed in S-2 and S-3 variety, similarly higher and
lower intensities were also observed in S-2 and S-3, respectively. Table 2 further shows
highest values of IV in S-2 variety and lowest IV in S-3, likewise IV was correlated with the
intensity of band at 1652 cm-1. It is well known fact that castor oil show high values of
viscosity. Among three varieties, highest viscosity was observed in S-2, while lowest in S-3
variety. These viscosity values highly correlated with the intensity of band at 1457 cm-1,
which showed similar trend. The band at 1377 cm-1 showed correlation with the values of
refractive index. The highest and lowest refractive index value was observed in S-3 and S-1,
similarly higher intensity was observed in S-2 and lower in S-1, respectively. PV also showed
correlation with two infrared bands at 1161 cm-1 and 1096 cm-1. As it can be seen from Table
2, that highest PV was observed in S-1 and lowest in S-3. Similar trend in higher and lower
intensity was observed in S-1 and S-3.

Table 5
FTIR spectral intensity of functional groups of castor seed oil
Frequency Intensity
S.No. Observation
(cm−1) S1 S2 S3
1 3421 0.0273 0.0279 0.0276 O─H stretching
C-H stretching vibration of the cis-
2 3007 0.0288 0.0293 0.0297
double bond (=CH)
3 2923 0.212 0.213 0.210 CH2 Asymmetrical stretching
4 2853 0.159 0.159 0.158 CH2 Symmetrical stretching
5 1742 0.232 0.233 0.221 C=O stretching
6 1652 0.0232 0.0240 0.0226 C=C Stretching
7 1457 0.0958 0.0972 0.0945 CH2 Scissors
8 1377 0.0714 0.0716 0.0718 Bending vibration of CH2 groups
9 1239 0.109 0.110 0.110
10 1161 0.193 0.192 0.188
Vibrations of the C─O ester groups
11 1096 0.136 0.135 0.131
12 1031 0.112 0.111 0.112
CH=CH (trans) bending out of
13 965 0.0784 0.0789 0.0789
plane
14 858 0.0765 0.0771 0.0777 =CH2 Wagging
Overlapping of the CH2 rocking
15 723 0.140 0.141 0.141 vibration and the out-of-plane
vibration of cis-disubstituted olefins

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Conclusion

The current investigation on indigenous local castor seed varieties encourages their
commercialization to increase economic growth at national level. A complete profile of local
varieties showed sufficient oil content (44–48%). Fatty acid examination by GCMS has
shown ricinoleic acid in high percentage. Some unusual fatty acids like isomers of oleic acid
and eicosenoic acid were also determined and reported in local varieties from this region.
FTIR study revealed the correlation of band intensities with physiochemical characteristics
of castor seed oil such as specific gravity, free fatty acids, saponification value, iodine value,
viscosity, refractive index and peroxide value.

Acknowledgments. The National Centre of Excellence in Analytical

Chemistry, University of Sindh, Jamshoro, Pakistan is gratefully
acknowledged for the financial support. The authors also would like to thank
Higher Education Commission (HEC), Pakistan.

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