MOJ Food Processing & Technology

Research Article Open Access

Biochemical studies on moringa oleifera seed oil

Abstract Volume 2 Issue 2 - 2016

The purpose of the present study was to examine the physico-chemical properties,
fatty acids composition, unsaponifiable matter, tocopherols and phenolic content Amany M Basuny, Maliha A Al-Marzouq
of Moringa oleifera seed oil from Al-Ahsa, Saudi Arabia. In addition, biological Food Science & Nutrition Department, King Faisal University,
evaluation of this oil was determined. Moringa oil showed a better overall quality, Saudi Arabia
its, acid, peroxide, iodine, saponification values. Sterol fraction was found rich in
β-sitosterol (45.11), stigmasterol (19.20%), campesterol (16.90%) and Δ5-avenasterol Correspondence: Amany M Basuny, Food Science & Nutrition
(10.00%). The major fatty acids were identified as oleic acid (65.00%). These results Department, Faculty of Agricultural Science & Foods, King Faisal
University, Saudi Arabia, Email dramany_basuny@yahoo.com
strongly suggested in potential use Moringa oil as non- conventional seed crop for
high quality oil. Received: November 3, 2015 | Published: February 24,
2016
Keywords: moringa oleifera, physico-chemical properties, fatty acids, biological
evaluation

Introduction The objectives of this study were to determine physico-chemical,

fatty acid composition, unsaponifiable matter and bioactive
Moringa oleifera lam. (M. olefera), a small deciduous tree, is components of oil extracted from the seeds of Moringa oleifera trees.
the most widely naturalized species of Moringaceae family and is In addition, biological evaluation of oil was evaluated.
commonly known as the horseradish or drumstick tree.1 It is native
in Asia Minor, Africa, the Indian subcontinent (Bangladesh, India & Materials and methods
Pakistan),2 and is also distributed in the Philippines, Cambodia, Central
America, North and South America, and the Caribbean Islands.1 The Plant materials
tree ranges in height from 5 to 12m and the fruits (pods) are around Moringa oleifera seeds (PKM1, variety) were procured from local
50cm long. When mature, the fruit of M. oleifera became and has market of Al-Ahsa, Saudi Arabia. The seeds were air-dried at room
10-50seeds inside. Fully mature dry seeds are round or triangular in temperature (25°C) for 1week.
shape and the kernel is surrounded by a light woody shell with three
papery wings.3 All parts of the Moringa tree-leaves, flowers fruits, Chemicals
and roots are edible and have long been consumed as vegetables4
All chemicals used were of analytical or HPLC grade from Merck
and used to treat many diseases such as abdominal tumors, hysteria,
(Darmstadt, Germany) or Sigma Aldrich (St. Louis, Mo, USA).
scurvy, paralytic attacks, helmintic bladder, prostate troubles, sores
Standards of sterols, tocopherol and phenolic compounds were
and skin infections.5 The leaves are highly nutritious, which contain
obtained from FlukaChemie (Buchs, Switzerland).
more vitamin A than carrots, more calcium than milk, more iron
than spinach, more vitamin C than oranges and more potassium than Oil extraction
bananas and more protein than milk and eggs.6 Moreover, leaves of
Moringa species are rich in various phytochemicals like carotenoids, The oil from the seeds was extracted with n-hexane using the
amino acids, sterols, glycosides, alkaloids, flavonoids, moringine, method described by Tsakins et al.14
moringinine, phytoestrogens, caffeoylquinic acid and phenolic
Proximate analysis
compounds.7 Fruits and seeds have been reported as a rich source of
protein, essential elements (Ca, Mg, K and Fe) and vitamins (A, C, The methods of the AOAC (2012) were used for proximate
and E). The oil extracted from its seeds (Known as ben or behen oil analysis. Moringa flower sample (5grams) was used for determination
due to the high behenic acid content) has a 38-40% yield and can of moisture content by weighing in crucible and drying in oven at
be used as a food, a cosmetic, and a lubricant.8 In African and some 105°C, until a constant weight was obtained. Determination of ash
parts of Asia, Particularly India, the oil has been used for cooking content was done by ashing at 550°C for 3hr. The kjeldah method was
purposes.9 In recent years, considering the gap between demand used to determine the protein content. The crude fiber content of the
and production of vegetable oils in many developing countries,9 samples was determined by digestion method and the fat was done by
research focusing on the use of unconventional oilseeds as a source Soxhlet extraction method. All determinations were done in triplicate.
of vegetable oils has become important. There are some reports on
the composition and characteristics of M. oleifera seed oil varieties Determination of the physico-chemical properties
from different countries of origin eg: India, Kenya, Malawi, Malaysia, The extracted Moringa oil was analyzed immediately for
Pakistan,3,7,10,11 considering its prospect as an alternative vegetable oil refractive index, color, acid value, peroxide value, iodine number and
source. The refined oil is clear, odorless and rancid-resistant. Seed saponification number as described in AOAC.15
biomass remaining after the oil extraction can be used as a fertilizer or
flocculating agent for water purification.12 The oil extracted from M. Oxidative stability
oleifera seeds is regarded as having a good commercial interest due to
The oxidative stability was estimated by measuring the oxidation
its physical, chemical and pharmacological characteristics.13
induction time, on a Rancimat apparatus (Metrohm CH series 679). Air

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© 2016 Basuny et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which
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Biochemical studies on moringa oleifera seed oil ©2016 Basuny et al. 41

(20L/h was bubbled through the oil (5.0g) heated at 100°C±2°C, with Biological evaluation
the volatile compounds being collected in water, and the increasing
water conductivity continually measured. The time taken to reach the For the biological evaluation, three types of diets were prepared:
conductivity inflection was recorded.16 a standard one, containing corn oil (control group), and the test diets
containing the Moringa oil and olive oil. The animals were divided
Fatty acid composition randomly in the groups of eight rats each: corn oil, Moringa oil and
olive oil. Diets (Table 1) were prepared according to AOCA15 and
Capillary gas chromatograph (HP 6890) was used for the qualitative stored under refrigeration (0-4°C) for not more than seven days.
and quantitative determinations of fatty acids of the oil and reported
in relative area percentages. Fatty acids were transesterfied into their Biochemical determinations
corresponding fatty acid methyl esters by shaking a solution of oil
(0.1g) in heptane (2ml) with solution methanolic potassium hydroxide After 49days, the animals were fasted overnight and anesthetized
(0.2ml, 2N). The fatty acid methyl esters were identified using a gas with ether for blood removal through the orbital plexus. Total
chromatograph equipped with DB-23 (5%-cyanopropyl-methyl poly cholesterol, HDL-cholesterols, triglycerides, albumin, total proteins,
siloxane) capillary column (60mx0.32mmX0.25µm film thickness) hemoglobin, trnasaminases, alkaline phosphatase (ALP), urea and
and flame ionization detector. Nitrogen flow rate was 0.6ml/min, creatinine were determined using the commercial kits (Labtest, Lagoa
hydrogen and air-flow rates were 45 and 450ml/min, respectively. The Santa, MG, Brazil).
oven temperature was isothermally heated 195°C. The injector and the Statistical analysis
detector temperatures were 230°C and 250°C, respectively. Fatty acid
methyl esters were identified by comparing their retention times with All experiments and measurements were carried out in triplicate,
known fatty acid standard mixture. Peak areas were automatically and the data were suggested to analysis of variance (ANOVA).
computed by an integrator. All GC measurements for each oil sample Analysis of variance and regression analyses were performed
were made in triplicate and the averages were reported. according to the MStat C and Excel software. Significant differences
between means were determined by Duncan’s multiple range tests. P
Identification of unsaponifiable matter values less than 0.05 were considered statistically significant.
The unsaponifiable matters of Moringa oil was analyzed by
an Hp 5890 gas chromatograph equipped with FID detector and
Results and discussion
DB-5 capillary column (30m, 0.25mm (5% phenyl)-95% methyl The results of the proximate composition of Moringa seeds are
polysiloxane, 0.25μm film thickness, 280°C temperature injector shows in Table 1. The moisture content of the Moringa seed was
and 300°C temperature transfer line. The oven temperature was 4.90%. The Moringa seed had higher values in the ash, crude fiber and
programmed as follows: initial temperature: 100°C for 2min, increase protein and carbohydrate contents. These values were higher than the
10°C/min up to 300°C, and then hold for 20min. The carrier gas values observed by Anwar et al.11 for Moringa flour. The high protein
was N2(2ml/min). The identification of the different compounds was content of these flour samples give an indication of their usefulness
performed by comparing of its relative retention times with those of in human diet and as livestock feed. The Moringa seed had higher
authentic reference compounds. fat content of 45.00%. The value was higher than the value (42%)
reported by Ogunsina et al.19
Determination of total phenolic content Table 1 Analysis of moringa oleifera seeds
The levels of total polyphenols of fresh crude sidr juice (fruit and
leaf) were determined according to the method of Gutfinger.17 Caffeic Constituents (%) M. oleifera seeds
acid was served as a standard compound for the preparation of the Moisture 4.90±0.55
calibration curve.
Oil 45.00±4.10
Tocopherol analysis
Protein 7.10±0.65
Tocopherol (α, β and δ) analysis was performed using an
Fiber 5.30±0.61
HPLC system consisting of a L-6000 Merck-Hitachi high pressure
ppm connected to an L-4000Merk Hitachi UV detector (Hitachi Ash 31.65±2.90
Instruments Inc., Tokyo, Japan) set at 295nm. Tocopherol contents Carbohydrate 10.95±1.15
were identified by comparing the retention times with those of pure
standards as described by others.7,18 AD-2500chromato Integrator Values are mean±SD calculated as percentage of dry weight for M. oleifera
(Merck, Darmstadt, Germany) was used for data acquisition and seeds, analyzed individually in triplicate.
processing. Table 2 shows various physico-chemical characteristics of the
extracted M. oleifera oil from Saudi Arabia. The values determined
Animals
for iodine number (69.01gI/100g oil), refractive index at 25°C
Twenty four day old male fisher rats were housed in individual (1.4570), saponification values (183.20mgKOH/g oil). However, the
cages and kept under controlled conditions of temperature and color at yellow 35.00 (2.00Red) value. Color of the oils is mainly
humidity with a light/dark cycle of 12-12hr. Diets and water were attributed to the presence of natural pigments which are extracted
offered ad libitum for 49days food consumption was monitored along with the oil during extraction and are effectively removed
weekly. during the bleaching step of processing of oil. The value of acidity
(0.60% as oleic acid) was considerably lower than M. oleifera oil
from India.20 Oils with lower values of acidity are more acceptable

Citation: Basuny AM, Al-Marzouq MA. Biochemical studies on moringa oleifera seed oil. MOJ Food Process Technol. 2016;2(2):40‒46.
DOI: 10.15406/mojfpt.2016.02.00030
 Copyright:
Biochemical studies on moringa oleifera seed oil ©2016 Basuny et al. 42

for edible applications. The peroxide value (0.83meq.kg-1 of oil) of clerosterol, 24-methylene cholesterol, Δ7-campestanol, Δ7-
which measure hydroperoxides of the oils, this value was lower than avenasterol, stigmastanol and 28. isoavenasterol, cholesterol and
those of M. oleifera oils from Kenya,21 and India.20 The induction brassicasterol. The contents of β-sitosterol, stigmasterol, campesterol
period (Rancimat:20L/h, 100°C±2°C) is an important feature which and Δ5-avenasterolin the present analysis of M. oleifera oil were rather
describes the oxidative stability of oil and fats.4 The induction period comparable with the values for M. Oleifera oil reported from Kenya
of the investigated M. oleifera oil (10.50h) was comparable with those and India. The sterol composition of the major constituents of the
M. oleifera oils reported from Sindh,4 but significantly lower than investigated M. oleifera oil generally varied to those of most of the
from India20 and Pakistan.22 conventional edible oils.24
Table 2 Physico-chemical properties of Moringa oleifera oil Table 4 Sterol composition of Moringa oleifera oil

Properties M. Oleifera oil Sterol (%) M. oleifera Oil

Refractive index (25°C) 1.4570±0.001
β-sitosterol 45.11±3.11
Color (Red unit) 2.00±0.33

Acidity (% as oleic acid) 0.60±0.09 Stigmasterol 19.20±1.33

Peroxide number (meq. O2/kg oil) 0.83±0.13

Campesterol 16.90±0.91
Iodine number (gI/100g oil) 69.01±5.30

Inductions period (hrs) 10.50±1.00 Δ5-avensterol 10.00±0.84

Values are mean±SD for M. oleifera oil, analyzed individually in triplicate. Clerosterol 1.20±0.19
Phenolic compounds have been proved to be responsible for
antioxidant activity on many vegetable seeds oils, it is mainly due to 24-methylene cholesterol 0.90±0.11
their redox properties, which can play an important role in absorbing
and neutralizing free radicals, quenching singlet and triplet oxygen or Δ7-campestanol 0.66±0.09
decomposing peroxides.23 Total phenolic compounds in the M. oleifera
seed oil was (160.00μg/g). These values were well comparable to Δ7-avenasterol 0.53±0.10
those reported in other Moringa species native to Kenya, Pakistan and
Egypt.
Stigmastanol 0.49±0.07
The data for tocopherol analysis of the M. oleifera oil from Saudi
Arabia are presented in Table 3. The levels of α, γ and δ-tocopherol 28-isoavenasterol 0.30±0.01
in the oil were 150.00, 70.80 and 55.50μg/g, respectively. The content
of α-tocopherol in the M. oleifera oil was in close agreement with the
Cholesterol 0.10±0.01
values reported for soybean, groundnut and palm oils.24 The contents
of γ-tocopherol in the M. oleifera oil was slightly lower than those
of M. oleifera oil from Malawi25 but considerably higher than those Brassicasterol 0.07±0.001
from India.20 The concentration of δ-tocopherol was slightly lower
than the values reported for M. oleifera oil from Kenya.21 Literature Values are mean±SD for M. oleifera oil, analyzed individually in triplicate.
revealed that α-isomer of tocopherol has greatest vitamin E potency, Table 5 shows the fatty acids composition in M. oleifera oil. This
whereas, δ-isomer of tocopherol has greater antioxidant efficacy than oil was found to contain a high level of oleic acid (65.00%), palmitic
either γ-or α-tocopherol.21,24 acid (12.31%) and linoleic acid (16.00%), palmitoleic acid (2.10%)
Table 3 Phenolic and tocopherol contents (ppm) of Moringa oleifera oil and stearic acid (5.10%), respectively. The content of principle fatty
acid, i.e. oleic acid was well in line with that reported for M. oleifera
Components M. oleifera Oil oil from Kenya21 and India.20 High oleic acid in Moringa oil makes
it desirable in the term of nutrition and high stability cooking and
Total phenolic compound 160.00±8.90 frying.3 The unsaturated fatty acids are very important for the stability
α-tocopherol 150.00±7.50 of oils because of the chemical reactions occurring at the double
bonds. The rate of those oxidation reactions depend on the number of
γ-tocopherol 70.80±4.10 double bonds in the carbon chain. Therefore, M. oleifera oil with high
δ-tocopherol 55.50±3.15
proportion of oleic acid is more stable than the others. In addition,
oleic is less susceptible to oxidation than polyunsaturated fatty acid
Values are mean±SD for M. oleifera oil, analyzed individually in triplicate. from the linoleic acid. Another interesting fact is that considerable
content of linoleic acid as an essential fatty acid in the M. oleifera oil
The sterols profile of M. oleifera oil is shown in Table 4. The may be provide high nutritional remuneration and render beneficial
sterol fraction of M. oleifera oil from Saudi Arabia mainly consisted healthy effect on blood lipid, blood pressure and cholesterol contents26
of β-sitosterol (45.11%), stigmasterol (19.20%), campesterol and it is preferred by industries when oil hydrogenation is required.
(16.90%) and Δ5-avenasterol (10.00%), together with small amounts

Citation: Basuny AM, Al-Marzouq MA. Biochemical studies on moringa oleifera seed oil. MOJ Food Process Technol. 2016;2(2):40‒46.
DOI: 10.15406/mojfpt.2016.02.00030
 Copyright:
Biochemical studies on moringa oleifera seed oil ©2016 Basuny et al. 43

Table 5 Fatty acid composition of Moringa oleifera oil Table 7 shows the sera total cholesterol contents of rats fed on M.
oleifera, olive and corn oils. The results indicate very little increases
Fatty acids M. oleifera oil (corn oil) and decreases (M. oleifera and olive oils). Changes in
C16:0 12.31±1.88
the levels of low-density lipoprotein cholesterol (LDL-C) of rats
administered M. oleifera, olive and corn oils are shown in Table 7.
C16:1 2.10±0.15 The results indicate that there were very little changes in the LDL-C
C18:0 5.10±0.80 levels of rats fed on diets containing investigated oils.

C18:1 65.00±5.13 Table 7 shows the changes in high-density lipoprotein cholesterol

(HDL-C) levels of rats fed on diets containing M. oleifera, olive and
C18:2 16.00±2.10 corn oils. The results demonstrate that the above-mentioned oils
C18:3 0.30±0.01 possessed very little change on the levels of HDL-C.

Values are mean±SD for M. oleifera oil, analyzed individually in triplicate. Table 8 shows the changes of urea and uric acid contents of rats
fed on M. oleifera, olive and corn oils. The results show that the
Table 6 shows sera AST activity of rats fed on M. oleifera oil, administration of oils induced very little change on the sera levels of
olive oil and corn oil. They were slight non-significant increase in urea and uric acid during the whole experiment.
the activity of AST during the whole experiment for rats fed on M.
oleifera seed oil, olive oil and corn oil. The data in Table 6 for rat The results of the present study were demonstrated that most of
sera activities of ALT and ALP on investigated oils indicate similar the characteristics and quality attributes of M. oleifera oil from Saudi
results for AST enzyme activity. Table 7 indicates the changes of total Arabia are quite identical with those of other Moringa oils reported in
lipid contents of rats administered M. oleifera, olive and corn oils. the literature. Its fatty acids composition revealed that it also fell in the
The results demonstrate that the administration of oil (M. oleifera and category of high oleic oils just like other Moringa oils. Also, contains
olive) used gradual significant decrease in the levels of total lipids. On high considerable amount of tocopherol and phenolic compounds
the contrary, the administration of corn oil caused significant increase similar to that found in olive oil and could be utilized in diet as a
in total lipid content of rat sera.27 source of vegetable oil human consumption.
Table 6 Influence of M. oleifera oil on the activity of serum AST, ALT, and AP (IU/L) of rats

Blood withdrawal period (week) Corn oil Olive oil M. oleifera oil
AST (IU/L)
0 40.10±2.33 40.10±2.33 40.10±2.33
1 40.23±1.98 40.63±2.54 40.31±1.99
2 40.71±2.46 40.61±2.78 40.45±2.01
3 40.75±2.86 40.60±2.60 40.66±2.75
4 40.33±1.87 40.76±2.98 40.15±1.79
ALT (IU/L)
0 44.00±3.11 44.00±3.11 44.00±3.11
1 44.15±3.23 44.20±3.25 44.16±3.56
2 44.80±3.81 44.36±3.67 44.40±3.39
3 44.85±3.80 44.76±3.51 44.81±3.78
4 44.46±3.25 44.52±3.49 44.64±3.54
AP (IU/L)
0 79.00±6.77 79.00±6.77 79.00±6.77
1 79.31±6.54 79.23±6.91 79.19±6.86
2 79.43±6.31 79.35±6.86 79.39±6.97
3 79.67±6.87 79.72±6.76 79.86±6.78
4 79.80±6.90 79.85±6.09 79.81±6.18

Values are mean±SD for M. oleifera oil, analyzed individually in triplicate.

Citation: Basuny AM, Al-Marzouq MA. Biochemical studies on moringa oleifera seed oil. MOJ Food Process Technol. 2016;2(2):40‒46.
DOI: 10.15406/mojfpt.2016.02.00030
 Copyright:
Biochemical studies on moringa oleifera seed oil ©2016 Basuny et al. 44

Table 7 Influence of M. oleifera oil on sera total lipid, total cholesterol, LDL-cholesterol and HDL-cholesterol (mg/dl) of rats

Blood withdrawal period (week) Corn oil Olive oil M. oleifera oil

Total lipids (mg/dl)

0 280.00±10.67 280.00±10.67 280.00±10.67

1 280.50±10.81 280.33±10.78 280.51±10.88

2 280.70±10.80 280.51±10.85 280.60±10.94

3 280.81±10.66 280.94±10.64 280.85±10.62

4 280.90±10.93 280.96±10.92 280.90±10.23

Total cholesterol (mg/dl)

0 160.00±7.98 160.00±7.98 160.00±7.98

1 160.81±8.01 160.85±8.16 160.90±8.12

2 160.55±7.99 160.70±8.45 160.85±8.41

3 160.60±8.25 160.65±7.56 160.93±8.43

4 160.66±8.33 160.80±8.09 160.95±8.21

LDL-cholesterol (mg/dl)

0 59.50±4.11 59.50±4.11 59.50±4.11

1 59.80±4.58 59.71±4.54 59.69±4.25

2 59.85±4.63 59.69±4.32 59.72±4.36

3 59.90±4.28 59.75±4.90 59.81±4.77

4 60.11±4.19 59.90±4.51 59.85±4.58

HDL-cholesterol (mg/dl)

0 100.10±6.01 100.10±6.01 100.10±6.01

1 100.15±6.12 100.20±6.58 100.33±6.38

2 100.25±6.45 100.35±6.42 100.50±6.78

3 100.30±6.66 100.45±6.23 100.61±6.25

4 100.60±6.39 100.50±6.89 100.81±6.75

Values are mean±SD for M. oleifera oil, analyzed individually in triplicate.

Citation: Basuny AM, Al-Marzouq MA. Biochemical studies on moringa oleifera seed oil. MOJ Food Process Technol. 2016;2(2):40‒46.
DOI: 10.15406/mojfpt.2016.02.00030
 Copyright:
Biochemical studies on moringa oleifera seed oil ©2016 Basuny et al. 45

Table 8 Influence of M. oleifera oil on the sera urea and uric acid levels (mg/dl) of rats

Blood withdrawal period (week) Corn oil Olive oil M. oleifera oil

Urea (mg/dl)

0 28.20±1.50 28.20±1.50 28.20±1.50

1 28.59±1.45 28.61±1.84 28.55±1.55

2 28.80±1.19 28.66±1.49 28.61±1.84

3 28.97±1.48 28.74±1.78 28.65±1.54

4 29.06±1.76 28.92±1.92 28.77±1.25

Uric acid (mg/dl)

0 4.10±0.33 4.10±0.33 4.10±0.33

1 4.30±0.39 4.51±0.53 4.20±0.74

2 4.41±0.45 4.55±0.57 4.51±0.54

3 4.50±0.40 4.60±0.65 4.76±0.38

4 4.66±0.56 4.67±0.39 4.81±0.85

Values are mean±SD for M. oleifera oil, analyzed individually in triplicate.

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DOI: 10.15406/mojfpt.2016.02.00030
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Citation: Basuny AM, Al-Marzouq MA. Biochemical studies on moringa oleifera seed oil. MOJ Food Process Technol. 2016;2(2):40‒46.
DOI: 10.15406/mojfpt.2016.02.00030