Industrial Crops and Products 82 (2016) 7480

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Industrial Crops and Products

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Solvent assisted extraction of oil from Moringa oleifera Lam. seeds

Payal R. Bhutada a , Ananda J. Jadhav b , Dipak V. Pinjari b, , Parag R. Nemade b ,
Ratnesh D. Jain b
a
Department of Oils, Oleochemicals and Surfactants, Institute of Chemical Technology, Matunga, Mumbai 400019, India
b
Department of Chemical Engineering, Institute of Chemical Technology, Matunga, Mumbai 400019, India

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

Article history: The present work reports the extraction of Moringa oil from the Moringao leifera Lam. (Moringaceae) seeds
Received 1 September 2015 using Soxhlet extraction. The effects of the various operating parameters such as temperature, solvent
Received in revised form ratio and solvent type on the extraction yield of the oil have been investigated. The extraction yield of
19 November 2015
the oil was varied with change in the operating temperatures (ranging from 80 to 110 C) as well as with
Accepted 4 December 2015
change in the solvents ratio (such as, chloroform:methanol (C:M) from 1:1, 2:1, 3:1, 4:1). The maximum
Available online 17 December 2015
extraction yield was obtained at C:M (3:1) ratio. The increase in the yield was also found with the increase
in the temperature. The fatty acid compositions of the Moringa oils extracted by the solvent extraction
Keywords:
Moringa oleifera Lam. seed
method have been analyzed using gas chromatography (GC). The change in the composition of the fatty
Moringa oil acid was seen with the change in the solvent media. Also, this extracted oil was analyzed by nuclear
Soxhlet extraction method magnetic resonance (NMR), Fourier transform infrared spectroscopy (FTIR), thermo-gravimetric analysis
(TGA) analysis. From TGA analysis it was found that, the oil degrade at temperature about 425450 C.
The oil was also analyzed with the physicochemical properties and the phytochemical tests to conrm
the presence of saponins, avonoids, steroids, terpenoids, phenols, and triterpenoids. It was found that
the solvent extraction is an effective method for the Moringa oil extraction.
2015 Elsevier B.V. All rights reserved.

1. Introduction
Moringa oleifera, known as Moringa, Drumstick, Malunggay is
an Indian tree grows in Asia, South America, Africa, the Caribbean
(Palafox et al., 2012). Moringa species is one of the most useful trees
in tropics and subtropics of Asia and Africa. Moringa belongs to the
Moringaceae family. Almost all parts of Moringa such as ower,
fruits, leaves, roots are edible and have been consumed as vegetable
(Abd El Baky Hanaa and El-Baroty Gamal, 2013). The root extract
possesses antimicrobial activity (Busani et al., 2012). The extracts
from the owers are found to have the hepatoprotective effect
(Upadhyay et al., 2015). The seeds have the edible oil known as
Ben oil. The seed cake is one of the best natural coagulants and can
be effectively utilized for treatment and purication of the highly
turbid water (Zhao and Zhang, 2013).
M. oleifera seeds contain 1947 % oil commercially known as
Ben oil and are rich in palmitic, stearic, behenic and oleic acids
(Ojiako and Okeke, 2013). Ben oil is reported to have very high
Corresponding author. Fax: +91 22 33611020.
E-mail addresses: dv.pinjari@ictmumbai.edu.in, dpinjari@gmail.com
(D.V. Pinjari).
oleic acid (70%) and smells a pleasant peanut like fragrance. Ben
oil is more stable than canola oil, soybean oil, palm oil when
used in frying (Nguyen et al., 2011). Ben oil has been used in
the manufacturing of perfume, hair care products, as lubricants,
for edible purpose. Ben oil is considered equivalent to olive oil
in terms of fatty acid composition (Zhao and Zhang, 2013). The
oil has fantastic cosmetic value; it is used in body and hair care
products as a moisturizer and as skin conditioner (Mahmood
et al., 2010). The oil consists of various sterols such as campes-
terol, stigmasterol, clerosterol, -sitosterol, 5 -avenasterol and less
quantity of 24-methylenecholesterol, campestanol, stigmastanol
and 28-isoavenasterol (Anwar et al., 2007). Another potential use
of Moringa oil is as a biodiesel feedstock, as an alternative source
of oil, Moringa seeds have been proposed as a potential source to
complement the mentioned feedstock (Palafox et al., 2012).
Moringa seeds and oil are used in the treatment of arthri-
tis, rheumatism, and hypertension (Aviara et al., 2015). Moringa
oil is very stable due to the presence of the high oleic acid
and so can be used for the frying application (Zhao and
Zhang, 2013). Moringa seed oil is non-drying oil and can be
used for lighting as it burns without dense smoke. Because of
the capacity of the edible oil to absorb and retain oral fra-
grances, it is valuable in perfume manufacturing. It is also used

http://dx.doi.org/10.1016/j.indcrop.2015.12.004
0926-6690/ 2015 Elsevier B.V. All rights reserved.
 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480 75

for lubricating watches and machinery. The avor of oil is some-

what similar to that of the peanut oil (Ghazali and Abdulkarim,
2011). Moringa seed oil has been widely used for the production
of biodiesel due to the high quantity of oil (around 40%). According
to the literature (Mojur et al., 2014a,b; Rashid et al., 2008), this
oil has excellent quality and can be used as raw material for the
biodiesel production (Atabani et al., 2014; Fernandes et al., 2015).
The most conspicuous property of biodiesel derived from M. oleifera
oil is the high cetane numbers of above 60, which are among the
highest reported for a biodiesel fuel (Kafuku et al., 2010).
Since the Egyptian times, Moringa oil has been used in skin
preparations and ointments (Mahmood et al., 2010). Moringa oil
is considered as a great cosmetic emollient. The oxidative stability
of the Moringa oil is due to the low content of the polyunsaturated
fatty acids. The oxidative stability of the oil is higher than that of
other oils which are rich in oleic acid (Ayerza (h), 2012). Due to its
various advantages and tremendous value, Moringa oil nds wide
application in cosmetic industry.
The solvent extraction technique is the oldest technique used for
the extraction of the oil. The solvent extraction technique has the
ability to perform the extractions on a liquid or a solid sample with
the minimal effort. Solvent extraction using chloroform:methanol
(C:M (1:1)) has been reported by (Lalas and Tsaknis, 2002). In the
present study, the solvent Soxhlet extraction of Moringa oil using
C:M ratios at various temperatures are reported. The objective of
this study is to develop the extraction process for recovering max-
imum oil yield from the Moringa seeds. In this work, comparative Fig. 1. Schematic of extraction set-up.

effect of oil extraction procedure on the physic-chemical charac-

teristics of oil extracted from M. oleifera Lam. seeds is studied. The
extraction yield and the fatty acid composition of oil extracted using
various solvent ratios at various temperatures will be compared.
2. Materials and methods
sample was once ltered through the lter paper and then the solu-
tion was collected and concentrated with the rotary evaporator to
acquire Moringa seed oil (Ramluckan et al., 2014; Tsaknis et al.,
1999a,b).

2.1. Samples and reagents

2.3. Yield determination

M. oleifera Lam. seed was purchased from local vendor Shamji

The extraction yield of the Moringa seed oil was calculated using
Amarshi & Sons, Herbal Departmental Stores, Mumbai. The Moringa
the following formula:
seed was cleaned to remove any foreign materials such as other
seeds, stones, etc. After cleaning the Moringa seed was soaked in M 
1
petroleum ether for 4 h to get rid of any other remained impurities. %Oil yield = 100 (1)
M2
After this, the seeds were further dried and then were crushed into
powder in a mixer. The powered seed was sieved from three sieve where M1 is the mass of Moringa seed oil extracted from the sample
of different mesh sizes (20, 40, 60 m). The powder from the sieve (g) and M2 is the mass of total material processed (g) by Soxhlet
of 40 m mesh size was used further for extraction. The resulted standard extraction.
seed powder was kept in lock bags until use. The reagents chloro-
form, methanol, petroleum ether used in extraction were analytical
grade and were purchased from SAF Scientic Chemicals Pvt., Ltd. 2.4. Energy calculation
Mumbai. All reagents were used as received without further puri-
cation. The energy required for the extraction of the Moringa seed oil
was calculated using the following equations:
2.2. Solvent extraction I
Power = voltage current (2)
The crushed, pre-dried Moringa seed powder (10 g) was placed P
in the paper thimble. The thimble is then placed into a Soxhlet
glass sample tube and sample tube was transferred to the extrac- Net energydelivered (J) = power (W) time(s) (3)
tion chamber in the Soxhlet apparatus (Fig. 1). A 200 ml of the
extraction solvent (ratios of chloroform:methanol) was transferred Quantity of material processed = solvent + seed powder (4)
into the solvent cup and placed on the heating plates. The cooling
water supply to the condensers was opened to ensure contin- J  
uous recycling of the solvent. The extraction chamber is made Net energy delivered
Net energy supplied =
in such a way that when the solvent surrounding the sample g Quantity of material processed
exceeds a certain level then it ows and trickles back down to (5)
the boiling ask. This cycle is repeated till the complete oil gets
extracted from the solid material. After this, the ask containing the
76 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480
3. Characterization of extracted oil
3.1. Physico-chemical characteristics of extracted oil
3.1.1. Determination of specic gravity and density
Specic gravity bottle (100 ml) was cleaned and dried in oven.
The weight of the empty gravity bottle was obtained as W1 , the
weight of the gravity bottle with the water was taken as W2 , and
the weight of the bottle with the oil was taken W3 . The specic
gravity and the relative density of the oil are calculated using the
formula below (Olushola, 2013):
 Weight of oil (W3 W1 )
Specic gravity =
Weight of equal volume of water (W2 W1 )
(6)
g
 Weight of oil 
Density( )=
ml Volume of oil
3.1.2. Determination of saponication value
2 g of extracted oil weighted and taken in conical ask and
approximately 25 ml 0.5 N alcoholic solution of potassium hydrox-
ide (KOH) was added. The mixture was then gently boiled under
reux on the water bath for 1 h. After this the titration of the hot
mixture was done by 0.5 N HCl, using phenolphthalein indicator.
The end point of the titration was faint pink (Adejumo et al., 2013).
 
28.05
Saponication value = (Blank-sample)
Weight of oil sample
3.1.3. Determination of free fatty acid
Acid value is expressed as % free fatty acid calculated as oleic
acid. 1 g of oil was weighted accurately and taken in the conical
ask. About 20 ml of neutral alcohol was added with few drops of
phenolphthalein and shaken vigorously. The solution was titrated
with 0.5 M alkali solution with constant shaking until pink color
remains constant. The acid value and the free fatty acid content
were calculated as follows: (Adejumo et al., 2013)
 0.5 56.1 Burette reading 
Acid value =
Weight of oil sample
 Acid value 
Free fatty acid =
2 In addition to the GC analysis, 1 H NMR was employed to study
3.1.4. Determination of iodine value
About 0.3 g of oil was taken in dry ask. Then 10 ml of car-
bon tetrachloride solution was added followed by 25 ml of WiJs
solution and was kept in the dark place for 30 min. Then 10% KI
solution was added in 20 ml to the mixture and was shaken. After
that, 100 ml of distilled water was and was shaken vigorously. Then
starch indicator was added to it and dark blue color was obtained.
The mixture was titrated with 0.1 N sodium thiosulphate till blue
to colorless point was obtained (Olushola, 2013). The iodine value
was calculated as follows:
 12.69

Iodine value = (Blank-sample) N
Weight of sample
3.2. Phytochemical screening
The phytochemical screenings were carried out to conrm the
presence or the absence of the secondary metabolitestannins,
saponins, avonoids, phenols, terpenoids, and steroids. Table 1
shows the details experimental process for determination of sec-
ondary metabolites (Ajibade et al., 2013).
3.3. Thermo-gravimetric analysis of Moringa oil
The thermo gram of extracted Moringa oil was recorded using
aluminum pan between temperature range 30500 C and under
inert atmosphere of N2 at a ow rate of 50 ml/min (Shimadzu,
Japan).
3.4. Gas chromatographic analysis of Moringa oil

Fatty acid composition of the Moringa seed oil extracted by
Soxhlet was determined using GC after derivatization to fatty acid
methyl esters (FAME). The preparation of the FAME was done by
standard IUPAC method. About 1 g of extracted oil was mixed with
0.1 g sodium methoxide and 10 ml of methanol and then the mix-
(7) ture was reuxed in water bath at 8090 C for about 1 h. The
mixture was cooled and taken in separating funnel with the addi-
tion of hexane followed by water washing. Finally, some sodium
sulphate was added to remove any traces of water in the FAME.
FAME separation and identication were carried out on a Shimadzu
gas chromatograph model GC-1000, equipped with BPX70 (70%
cyanopropyl polysilphenylene-siloxane) highly polar capillary col-
umn and ame ionized detector. The amount of the sample injected
was 1.0 l. Nitrogen ow rate of 4.0 ml/min was used as the car-
rier gas and a split injector was used with a split ratio of 50:1. The
injector and detector temperatures were set at 270 C and 280 C
respectively. The oven temperature was programmed as follow:
the initial temperature was 180 C which was maintained for 4 min
(8) from 180 C to 190 C at 2 C/min, from 190 C to 270 C at 3 C/min
hold for 6 min, and then to a nal temperature 320 C hold for
another 6 min. Fatty acid methyl esters were quantied as per-
centages of the total methyl ester peak areas (Bezerra and Filho,
2014).
3.5. FTIR analysis of Moringa oil
FTIR analysis was used to reveal the functional groups present
in the extracted oil. The FTIR spectra of extracted oil samples was
recorded using FTIR spectrophotometer (Shimadzu 8400s, Japan)
using ATR sampling technique by recording 45 scan in % transmit-
tance mode in the range of 4000500 cm1 .
(9)
1H
3.6. NMR analysis of Moringa oil
(10)
and compare the molecular structure of the Moringa seed oil
extracted using the solvent extraction technique. 1 H NMR of the
extracted Moringa oil was analyzed using Agilent 400 MHz. The oil
sample was dissolved in the deuterated chloroform and the spectra
were recorded (Almoselhy et al., 2014).
4. Results and discussion
4.1. Effect of temperature
As shown in the Table 2, the yield of the Moringa seed oil changes
with the increasing temperature in the solvent extraction. When
(11)
temperature increases (gradually) from 70 C to 100 C, the extrac-
tion yield was increases by about 10% for C:M (2:1) solvent ratio
and by about 10% for C:M (3:1) solvent ratio. Fig. 2(A) shows the
effect of temperature on the extraction yield of Moringa oil for
the solvent ratio of C:M (2:1). As the temperature increases from
70 C to 100 C extraction of oil was increased due to the increase
of solubility and diffusivity which improves the mass transfer.
 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480 77

Table 1
Detailed description of experimental procedure for conrmation of presence of secondary metabolites.

Metabolites Experimental procedure

Tannins 0.5 g of the moringa oil was mixed with 20 ml water. The mixture was boiled and ltered. Then few drops of 0.1% ferric
chloride were added. The development of the brownish green or blueblack color conrms the presence of tannins.
Saponins 2 g of moringa oil was mixed and boiled with 20 ml of the water and then ltered. 10 ml of this ltrate was further
mixed with 5 ml of distilled water and was shaken vigorously for stable persistent froth. The formation of the froth
conrms the presence of the saponins.
Flavonoids The known quantity of the oil was taken with the ammonia solution and the concentrated sulfuric acid was added and
was allowed to develop the color. Development of yellow color indicated the presence of the avonoids.
Steroids 0.5 g of the moringa oil was mixed with the 2 ml of acetic anhydride with further addition of concentrated sulfuric
acid and was allowed to develop color. The color change from violet to blue or green color conrms the presence of
the steroids.
Phenols 1 g of the oil was mixed with 20 ml water with the further addition of the ferric chloride. The appearance of the blue
color indicated the presence of the phenol.
Terpenoids 5 g of the oil was mixed with 2 ml of the chloroform. Further 3 ml of concentrated sulphuric acid was added to form
the layer. Development of the reddish brown color at interface indicates presence of the terpenoids.

Table 2
Comparison of extraction results with % yield and energy supplied.

Exp. Run Solvent Temp. ( C) Time (min.) Avg.% yield Std. Dev. Net energy supplied (kJ/g)

E1 C:M (1:1) 80 180 17.8 5.37 11.83

E2 C:M (2:1) 70 240 28.8 1.13 16.91
E3 C:M (2:1) 80 120 31.0 2.83 7.885
E4 C:M (2:1) 100 90 38.0 4.24 6.34
E5 C:M (2:1) 110 90 34.4 0.57 6.34
E6 C:M (3:1) 80 60 31.5 6.36 4.532
E7 C:M (3:1) 100 69 41.0 0.04 5.211
E8 C:M (4:1) 80 120 37.0 1.41 9.403
E9 C:M (4:1) 100 120 36.0 1.41 9.49
E10 Methanol 80 210 11.8 1.1 20.7
E11 Chloroform 80 60 24.2 4.53 3.247
E12* Petroleum ether 80 360 28.6 1.41 38.96
E13 C:M (1:1) 80 360 21.1 0.57 23.66
E14 Petroleum ether 100 120 0.0 0.00 15.5
E15 C:PE:M(75:5:20) 80 150 26.6 2.26 11.7
*
Raw powder was rst extracted in E12 experimental run and the spent powder was used in next E13 experimental run for extraction with C:M.

Fig. 2. (A) Effect of temperature on extraction yield for C:M (2:1); (B) effect of solvent typeon the extraction yield of moringa oil; (C) effect of solvent ratio on the extraction
yield of the Moringa oil.
78 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480
Table 3
Physicochemical characteristics of extracted moringa oil.
Constituents Obtained FAO/WHO standard
Density (gm/cm3 ) 0.24 0.009
Specic gravity 0.98 0.006 0.9
Acid value (mg KOH/gm) 26.22 1.210 4 The test for the presence of the avonoids, saponins, phenol,
Free fatty acid (mg KOH/gm) 13.11 0.615 5.77.28
Saponication value (mg KOH/gm) 172.16 0.945 181.4
Iodine value 75.06 0.943 80106
However, when temperature exceed beyond the 100 C the yield
decreases. The possible reason behind this nature may be the reduc-
tion and breakdown of antioxidant molecule at high temperature.
Another possibility may be the decomposition of active ingredi-
ents presents in the oil (Hossain et al., 2009). Also, from Table 2 it
is clear that, as the temperature increase, the yield of the Moringa
oil increases and the energy supplied per unit of extracted oil was
decreases. The highest extraction yield of oil (41%) was obtained
at the solvent ratio of 3:1 (C:M) and temperature of 100 C with
expense of energy requirement of 5.211 kJ/g of oil.
4.2. Effect of solvent type
The solvents used for the extraction in this work, were both polar
(methanol) and non-polar (chloroform, petroleum ether). Polarity
is dened as the relative ability of the molecule to engage in the
strong interactions with the other polar molecules. Polarity rep-
resents the ability of the molecule to enter into interactions of
all kinds. Relative polarity is the sum of all possible interactions
(Hossain et al., 2009). It was found that, as the polarity of the solvent
decreases, the extraction yield oil increases as shown in Fig. 2(B).
The yield for the methanol is low as compared to the yield for the
chloroform and petroleum ether as shown in Table 2. Polar solvents
are highly capable of extracting the nonfat material such as antiox-
idants, tocopherol, pigments, etc., non polar solvents along with
the non-fatty matter also extract the oil present in the seeds. The
oil extracted using only methanol was red in color whereas the oil
extracted using the chloroform and the petroleum ether was yel-
low in color. The red color of the oil may be due to the presence of
the pigments and the tocopherol present in the Moringa seed.
4.3. Effect of solvent ratio
The extraction of the Moringa oil was performed using the sol-
vent mixture of chloroform and methanol in various ratios. The
comparison of the extraction of Moringa oil with respect to various
solvent ratios is shown in Fig. 2(C). The solvent mixture contains
both the polar (methanol) and the nonpolar (chloroform) solvent.
The ratio of the nonpolar solvent was changed and the difference
in the extraction yield of the oil was noted. With the increase in
the nonpolar solvent from 1:1 to 3:1 the yield in the extraction of
the oil also increases. However, when the ratio of nonpolar solvent
in the mixture was increased further then the yield was depressed.
This is because, nonpolar solvent may be unable to extract the toco-
pherols, pigments, antioxidants and the other substances present
in the oil.
4.4. Physico-chemical analysis of extracted Moringa oil
The physicochemical properties such as density, specic grav-
ity, acid value, saponication value, iodine value, were analyzed
and are tabulated in Table 3. The specic gravity, iodine value and
saponication value of the extracted oil matches with the reported
values (Table 3). The acid value and free fatty acid content was sig-
nicantly higher than that of the reported values and it will vary as
per the geographical locations where Moringa plants are grown.
4.5. Phytochemical analysis of extracted Moringa oil
tannins, and terpenoids were done and found to be positive. These
chemical constituents are known for many therapeutic values and
various applications will be determined on the basis constituents
present in the oil. The avonoid test showed the presence of yellow
color, the froth formation was seen in the saponins test, reddish
brown color indicated the presence of the terpenoids, and phenols
test gave the light red color. Grey color was seen in the tannins test
and red color for the steroid test. The avonoids are reported to have
the antifungal, antibacterial and antioxidant properties (Emmanuel
et al., 2014).
4.6. Gas chromatography (GC) analysis of extracted Moringa oil
Fig. 3(A) shows the GC chromatogram of extracted oil which
conrms the presence of the various fatty acids such as palmitic,
stearic, arachidic and behenic acids and the main unsaturated
fatty acid is oleic acid (70.5%), with small amounts of palmitoleic,
linolenic, and eicosenoic acids (Table 4). Since, the extracted oil
contained a substantial amount of behenic acid (4.9%), it can be
used as a natural source of behenic acid, which has been used as an
oil structuring and solidifying agent in margarine, shortening, and
foods containing semi-solid and solid fats, eliminating the need to
hydrogenate the oil (Abdulkarim et al., 2005). The high percentage
of oleic acid in the oil makes it desirable in terms of nutrition and
high stability cooking and frying oil. High oleic-acid vegetable oils
such as high-oleic corn, sunower and canola have been found to
have enough oxidative stability to be used in demanding applica-
tions such as frying (Petukhov et al., 1999; Warner and Knowlton,
1997). In addition high-oleic oils have low saturated fatty acid lev-
els. Therefore high-oleic oils can be viewed as a healthy alternative
to partially hydrogenated vegetable oils.
4.7. FTIR analysis of extracted Moringa oil
FTIR analysis as shown in Fig. 3(B) clearly shows one peak
of ester functional group (C O stretch) at 1773 and peak at
1162 shows ether linkage and broad peak of 3414 (O H stretch)
and 750 peaks of ortho-substituents. The peak in the region
35003200 cm1 indicates the presence of the alcohol or phenol
(O H stretch). The elongated U shape around this region shows
the presence of the alcohol group. This functional group appears
predominantly in the protein and fatty acid structures present
in Moringa seeds. Due to the high content of protein present in
seeds there is also a contribution in this region from the N H
stretching of amide groups. The peaks at 2920 cm1 and 2852 cm1
are assigned to symmetrical and asymmetrical stretching of the
C H of CH2 group present in fatty acids. One peak in the region
17601665 cm1 shows the presence of the ester functional group.
The peak in the region 30002700 cm1 shows the presence of the
carboxylic acid. The peak in the region of 15001450 cm1 shows
the presence of the benzene (C C) (Arajo et al., 2010).
4.8. NMR analysis of extracted Moringa oil
It was observed from the 1 H NMR analysis Fig. 3(C), there were
no signals for hydroperoxides (primary oxidation products) and
aldehydes (main secondary oxidation products), indicating that no
oxidative degradation was taking place. The presence of these oxi-
dation products in the ranges 8.098.19 ppm for hydroperoxides
proton, and 9.309.90 ppm for aldehydes, indicates oxidation of
 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480 79

Fig. 3. (A) Gas chromatic analysis of Moringa oil; (B) FTIR Spectra of Moringa oil; (C) 1 H NMR spectra of oil extracted by solvent extraction; (D) TGA analysis of Moringa oil.

Table 4
Relative percent composition of fatty acid in Moringa oleifera seed oil.

Fatty acids Determined values Reported values

Abdulkarim et al. (2005) Sunga and Whitby (1995) Dahot and Memon (1985) Ferrao and Ferrao (1970)

(C16:1) 2.2 2.2 1.1

(C18:0) 7.7 7.6 8.3 8.3 4.3
(C18:1) 73.5 67.9 67.7 67.3 76.5
(C20:0) 4.6 4 4.7 2.7 2.7
(C20:1) 2.9 1.5 2.3
(C22:0) 4.9 6.2 7.4 5.6 4.6
(C24:0) 1 6.2 0.4 3.2 1.1

these edible oils (Alonso-Salces et al., 2011). The presence of sterol
(-sitosterol) in extracted oil could be deduced from the signal
0.84 ppm due to methylic protons in position C18 of the steroidal
skeleton (Almoselhy et al., 2014). The peak in the region 0.840.85
(A) is the methyl proton, splitted into triplets. The acyl group in
the region 1.121.16 (B) has been splitted into multiplets. Other
aliphatic protons are splitted in the region of 1.572.29 (CE). The
alcoholic OH shows triplet at 3.433.44 (F) (downeld shifting of
protons of OH due to electro negativity of oxygen). Esteric proton
shows peak at 3.62 (G). Glyceryl group shows peak at 4.084.13
(H). The CH2 O of tetrahydropyran ring shows peak at 4.254.28
(I). The CH2 O between tetrahydropyran ring ( O ) and ester group
shows peak at 5.3 (J) (more downeld because of electronegative
group on both sides). The solvent (deuterated chloroform) shows
peak at 7.3 (K).
4.9. TGA analysis of extracted Moringa oil
The thermo-gravimetric analysis of the extracted Moringa oil
was carried out to characterize the decomposition stages and
thermal stability of the Moringa oil as shown in Fig. 3(D). This
thermo gravimetric curve veries the sample heterogeneity, since
80 P.R. Bhutada et al. / Industrial Crops and Products 82 (2016) 7480
the intermediates formed are a mixture of several components.
The mass loss verses temperature curve has been divided into
three stages. First stage from 30 C to 130 C having mass loss of
10%, associated with water desorption. In the second step remain-
ing 40% of mass loss was observed in the temperature range of
130300 C. This stage occurs due to the decomposition of organic
matter, probably the protein components, present in Moringa oil.
The third stage shows the remaining 40% mass loss occurs from
300 C to 428 C with decomposition of the greater part of the oil
components, which probably includes fatty acids, for example oleic
acid has a boiling point of 360 C.
5. Conclusions
The solvent extraction process for extracting oil from M. oleifera
seeds was successfully developed after evaluating and comparing
with various solvent ratios at various temperatures. The extracted
oils were liquid and stable at room temperature. The oil extraction
using C:M (3:1) solvent ratio at 100 C gave the highest yield. The
extraction of oil using various solvent is less as compared to extrac-
tion of oil using mixture of solvents at various ratios and the yield
changes with the changing solvent ratios and temperatures. The
solvent ratios were used to extract oil as well as other materials
such as antioxidants, tocopherol, pigments, sterols, etc., present in
the seeds. The polar solvent was mainly responsible for increasing
the yield of the oil by extracting the other materials.
The GC analysis showed the fatty acid composition of the
extracted Moringa oil. The other analysis technique such as FTIR
and 1 H NMR conrmed the functional group and the structure of
components presents in the extracted oil. TGA analysis was done for
the extracted Moringa oil to see the weight loss with respect to the
temperature. The initial and nal weight loss showed the presence
of the aliphatic and the aromatic segments. Finally, it is conrmed
that, the solvent extraction technique using Soxhlet apparatus is
an effective and indeed feasible method for the extraction of the
Moringa seed oil.
Acknowledgement
The co-authors (D. V. Pinjari and A. J. Jadhav) would like to
acknowledge Department of Science and Technology, Government
of India for their continuous support for carrying out the proposed
research work. Also another co-author (P. R. Nemade) would also
like to thank UGC for providing fund under UGC-FRPS (Faculty
Research Promotion Scheme) for this project.
References
Abdulkarim, S.M., Long, K., Lai, O.M., Muhammad, S.K.S., Ghazali, H.M., 2005. Some
physico-chemical properties of Moringa oleifera seed oil extracted using
solvent and aqueous enzymatic methods. Food Chem. 93, 253263.
Abd El Baky Hanaa, H., El-Baroty Gamal, S., 2013. Characterization of Egyptian
Moringa peregrine seed oil and its bioactivities. Int. J. Econ. Bus. Res. 2, 99108.
Adejumo, B.A., Alakowe, A.T., Obi, D.E., 2013. Effect of heat treatment on the
characteristics and oil yield of Moringa oleifera seeds. Int. J. Eng. Sci. 2, 223239.
Ajibade, T.O., Arowolo, R., Olayemi, F.O., 2013. Phytochemical screening and
toxicity studies on the methanol extract of the seeds of Moringa oleifera. J.
Complement. Integr. Med. 10.
Almoselhy, R.I.M., Allam, M.H., El-Kalyoubi, M.H., El-Sharkawy, A.A., 2014. H NMR
spectral analysis as a new aspect to evaluate the stability of some edible oils.
Ann. Agric. Sci. 59, 201206.
Alonso-Salces, R.M., Holland, M.V., Guillou, C., 2011. 1 H NMR ngerprinting to
evaluate the stability of olive oil. Food Control 22, 20412046.
Anwar, F., Latif, S., Ashraf, M., Gilani, A.H., 2007. Moringa oleifera: a food plant with
multiple medicinal uses. Phytother. Res. PTR 21, 1725.
Arajo, C.S.T., Alves, V.N., Rezende, H.C., Almeida, I.L.S., de Assunco, R.M.N., Tarley,
C.R.T., Segatelli, M.G., Coelho, N.M.M., 2010. Characterization and use of
Moringa oleifera seeds as biosorbent for removing metal ions from aqueous
efuents. Water Sci. Technol. J. Int. Assoc. Water Pollut. Res. 62, 21982203.
Atabani, A.E., Mojur, M., Masjuki, H.H., Badruddin, I.A., Kalam, M.A., Chong, W.T.,
2014. Effect of Croton megalocarpus, Calophyllum inophyllum, Moringa oleifera,
palm and coconut biodieseldiesel blending on their physico-chemical
properties. Ind. Crops Prod. 60, 130137.
Aviara, N.A., Musa, W.B., Owolarafe, O.K., Ogunsina, B.S., Oluwole, F.A., 2015. Effect
of processing conditions on oil point pressure of Moringa oleifera seed. J. Food
Sci. Technol. 52, 44994506.
Ayerza (h), R., 2012. Seed and oil yields of Moringa oleifera variety Periyakalum-1
introduced for oil production in four ecosystems of South America. Ind. Crops
Prod. 36, 7073.
Bezerra, K. da S., Filho, N.R.A., 2014. Characterization and quantication by gas
chromatography of free steroids in unsaponiable matter of vegetable oils. J.
Braz. Chem. Soc. 25, 238245.
Busani, Moyo, Patrick Julius, Masika, Muchenje, V., 2012. Antimicrobial activity of
Moringa oleifera (Lam.) root extract. Afr. J. Biotechnol. 11, 27972802.
Dahot, M.U., Memon, A.R., 1985. Nutritive signicance of oil extracted from
Moringa oleifera seeds. J. Pharmacol. Univ. Karachi 20, 7579.
Emmanuel, S., Emmanuel, S.A., Olajide, O.O., Abubakar, S., Idowu, I.D., Orishadipe,
A.T., Thomas, S.A., 2014. Phytochemical and antimicrobial studies of methanol,
ethyl acetate, and aqueous extracts of Moringa oleifera seeds. Am. J. Ethnomed.
1, 346354.
Fernandes, D.M., Sousa, R.M.F., de Oliveira, A., Morais, S.A.L., Richter, E.M., Munoz,
R.A.A., 2015. Moringa oleifera: a potential source for production of biodiesel
and antioxidant additives. Fuel 146, 7580.
Ferrao, A.M.B., Ferrao, J.E.M., 1970. Acidos gordos em oleo de moringuerio. Agron.
Angolana 30, 316.
Ghazali, H.M., Abdulkarim, S.M., 2011. In Moringa Oleifera Seed Oil: Composition,
Nutritional Aspects and Health Attributes. Elsevier Life Sciences, pp. 787794.
Hossain, M.A., A.L-Mijizy, Z.H., Al-Rashdi, K.K., Weli, A.M., Al-Riyami, Q., 2009.
Effect of temperature and extraction process on antioxidant activity of various
leaves crude extracts of Thymus vulgaris. J. Coast. Life Med. 1, 130134.
Kafuku, G., Lam, M.K., Kansedo, J., Lee, K.T., Mbarawa, M., 2010. Heterogeneous
catalyzed biodiesel production from Moringa oleifera oil. Fuel Process. Technol.
91, 15251529.
Lalas, S., Tsaknis, J., 2002. Characterization of Moringa oleifera seed oil variety
periyakulam 1. J. Food Compos. Anal. 15, 6577.
Mahmood, K.T., Mugal, T., Haq, I.U., 2010. Moringa oleifera: a natural gifta review.
J. Pharm. Sci. Res. 2, 775781.
Mojur, M., Masjuki, H.H., Kalam, M.A., Atabani, A.E., Arbab, M.I., Cheng, S.F., Gouk,
S.W., 2014a. Properties and use of Moringa oleifera biodiesel and diesel fuel
blends in a multi-cylinder diesel engine. Energy Convers. Manage. 82, 169176.
Mojur, M., Masjuki, H.H., Kalam, M.A., Atabani, A.E., Fattah, I.M.R., Mobarak, H.M.,
2014b. Comparative evaluation of performance and emission characteristics of
Moringa oleifera and palm oil based biodiesel in a diesel engine. Ind. Crops
Prod. 53, 7884.
Nguyen, H.N., Gaspillo, P.D., Maridable, J.B., Malaluan, R.M., Hinode, H., Salim, C.,
Huynh, H.K.P., 2011. Extraction of oil from Moringa oleifera kernels using
supercritical carbon dioxide with ethanol for pretreatment: optimization of
the extraction process. Chem. Eng. Process. Process Intensif. 50, 12071213.
Ojiako, E.N., Okeke, C.C., 2013. Determination of antioxidant of Moringa oleifera
seed oil and its use in the production of a body cream. Asian J. Plant Sci. Res. 3,
14.
Olushola, S. musliu, 2013. Extraction and characterization of Moringa oleifera seed
oil. Res. Rev. J. Food Dairy Technol. 1, 2227.
Palafox, J.O., Navarrete, A., Sacramento-Rivero, J.C., Rubio-Atoche, C., Escofe, P.A.,
Rocha-Uribe, J.A., 2012. Extraction and characterization of oil from Moringa
oleifera using supercritical CO2 and traditional solvents. Am. J. Anal. Chem. 03,
946949.
Petukhov, I., Malcolmson, L.J., Przybylski, R., Armstrong, L., 1999. Frying
performance of genetically modied canola oils. J. Am. Oil Chem. Soc. 76,
627632.
Ramluckan, K., Moodley, K.G., Bux, F., 2014. An evaluation of the efcacy of using
selected solvents for the extraction of lipids from algal biomass by the soxhlet
extraction method. Fuel 116, 103108.
Rashid, U., Anwar, F., Moser, B.R., Knothe, G., 2008. Moringa oleifera oil: a possible
source of biodiesel. Bioresour. Technol. 99, 81758179.
Sunga, I., Whitby, G., 1995. Decentralised Edible Oil Milling in Zimbabwe: an
Evaluation Report of the Tinytech Oil Mill Project. In Progress Report for
Intermediate Technology Development Group, (Intermediate Technology).
Tsaknis, J., Spiliotis, V., Lalas, S., Gergis, V., Dourtoglou, V., 1999a. Quality changes
of Moringa olifera, variety Mbololo of Kenya, seed oil during frying. Grasas
Aceites 50.
Tsaknis, J., Lalas, S., Gergis, V., Dourtoglou, V., Spiliotis, V., 1999b. Characterization
of Moringa oleifera variety Mbololo seed oil of Kenya. J. Agric. Food Chem. 47,
44954499.
Upadhyay, P., Yadav, M.K., Mishra, S., Sharma, P., Purohit, S., 2015. Moringa oleifera:
a review of the medical evidence for its nutritional and pharmacological
properties. Int. J. Res. Pharm. Sci. 5, 1216.
Warner, K., Knowlton, S., 1997. Frying quality and oxidative stability of high-oleic
corn oils. J. Am. Oil Chem. Soc. 74, 13171322.
Zhao, S., Zhang, D., 2013. A parametric study of supercritical carbon dioxide
extraction of oil from Moringa oleifera seeds using a response surface
methodology. Sep. Purif. Technol. 113, 917.