Groudnut Oil

INTERNATIONAL JOURNAL OF SCIENTIFIC PROGRESS AND RESEARCH (IJSPR) ISSN: 2349-4689
Issue 168, Volume 68, Number 01, February 2020
Extraction of Edible Oil From Groundnut By using

Solvents and Enzymes
B. Samatha1, Sevya Naik2
1,2
Department of Chemical Engineering, University College of Technology, Osmania University, Hyderabad, Telangana, India.
Abstract - The methods of oil aqueous extraction process (AEP) assisted by enzymes are, over the last 50 years, an alternative designed
to replace traditional methods of extraction using organic solvents. To extract the oil using an AEP, the use of specific enzymes, able
to hydrolyze some or all components of seeds can significantly increase the yields of extraction. Hydrolyzing the different constituents
of cell walls (cellulose, hemicellulose, pectin, proteins, etc.), enzymes is able to enhance the liberation of the oil. A number of physico-
chemical parameters must also be considered for the better expression of the enzymatic mixture, while maintaining the quality of oils
and meals. An experimental study was performed in order to investigate the effect of an enzymatic pre-treatment process for extraction
of oil from groundnuts. In the present study celluclast 1.5L was used for the pre-treatment. The effect of enzyme concentration (5-
10%), temperature (50-60⁰C), pH (5.0- 6.0), reaction time (1-7 h) on free oil liberated was studied. Residual oil was collected by
subjecting the treated meal to soxhlet extraction for 4 h. The optimal conditions were: enzyme concentration of 7.5% (w/w) in 10 g of
peanut seeds, pH 5.0, 50°C, and 5 h with constant shaking at 450 rpm. Centrifuging the mixture at 8500 for 20 min separated the oil
with a recovery of 71-73.1%.
Keywords: Extraction; ediableoil; solvents; enzymes.
I. INTRODUCTION
„Lipids‟ are fatty acids and their derivatives, and substances related biosynthetically or functionally to these compounds.
These are insoluble in water but soluble in organic solvents such as chloroform, hydrocarbons, alcohols, esters. Lipid is a
chemical term being used for different types of compounds like triacylglycerol. They are relatively simple molecules, for
example the fatty acid they are more complex and contain phosphor or sulpho groups, amino acid, peptides and their
derivatives. Lipids can be classified as derived, simple, or complex. The derived lipids include fatty acids and alcohols,
which are building blocks for simple and complex lipids. Simple lipids composed of fatty acids and alcohols include
acylglycerols, ether acylglycerols, sterols and their esters and wax esters. Complex lipids include glycerophospholipids,
glyceroglycolipids and sphingolipids. These structures yield three or more different compounds on hydrolysis. Particularly
common and practically useful system is the division in to neutral lipids and polar or amphiphilic lipids.
Thus neutral lipids include simple hydrocarbons, carotenes, triacylglycerol‟s, wax, esters, and sterol esters.Other lipids
such as fatty acids, polyprenols, sterols in which hydrophilic function has relatively little impact on the overall molecular
characterization. However, it is evident that oils and fats are one of the major components of lipids. Oils and fats are
predominantly tri esters of fatty acids and glycerol, commonly called „triglycerides or triacylglycerol‟s‟. Substances which
are solid or semisolid at room temperature are called „fats‟ and those which are liquids are called „oils‟.
Fatty Acids
Fatty acids are the building blocks for triacylglycerol. Aliphatic carboxylic acids with 4 or more carbon atoms are called
fatty acids. In nature, they occur with an even number of carbon atoms, with very few exceptions. The natural oils and fats
contain saturated and unsaturated fatty acid. Saturated means that all the carbon valences (except in the carboxylic acid
group) are satisfied. Fatty acids having hydrogen deficient carbon atoms bonded by double or triple valences are called
unsaturated. According to the number of double bonds present they are called as mono unsaturated or di-, tri-, tetra-, penta,
hexa-enoic acids etc. If the fatty acids contain more than one double bond, those are called as poly unsaturated fatty acids
(PUFA).
The most common fatty acids present in the edible oils are lauric, myristic, palmitic (saturated), oleic (mono unsaturated),
linoleic and linolenic (poly unsaturated) acids and the classification is shown in Fig1.
www.ijspr.com IJSPR | 1
Fig 2: Structure of Cisand Trans-Isomers

However, some oils contain unusual fatty acids like hydroxy fatty acids (ricinoleic acid in castor oil) also. Other fatty acids
are arachidonic (AA), eicosapentaenoic (EPA) and docosahexanoic (DHA) acids present in fish oil. Examples of few
common unsaturated and saturated fatty acids are tabulated below in Table 1 and 2.
Most of the hydrogen in double bonds of natural fatty acids is found on the same side of the bond, indicating a
„Cis position‟ and the respective structures are shown in Fig 2. But, some of the hydrogen atoms may move to the other side
of the bond during dehydrogenation process, to produce „Trans-isomers‟.
The properties of the fatty acids which depend up on their degree of saturation or unsaturation with respect to hydrogen are
retained by them when they are in combination with glycerin as triglycerides.
Table 1: Saturated fatty acids
Common name Chemical structure C:D
Caprylic acid CH3(CH2)6COOH 8:0
Capric acid CH3(CH2)8COOH 10:0
Lauric acid CH3(CH2)10COOH 12:0
Myristic acid CH3(CH2)12COOH 14:0
Palmitic acid CH3(CH2)14COOH 16:0
Stearic acid CH3(CH2)16COOH 18:0
Arachidic acid CH3(CH2)18COOH 20:0
Behenic acid CH3(CH2)20COOH 22:0
Lignoceric acid CH3(CH2)22COOH 24:0
Cerotic acid CH3(CH2)24COOH 26:0
Table 2: Unsaturated fatty acids
Common name Chemical structure Δx C:D n−x
Myristoleic acid CH3(CH2)3CH=CH(CH2)7COOH cis-Δ9 14:1 n−5
Palmitoleic acid CH3(CH2)5CH=CH(CH2)7COOH cis-Δ9 16:1 n−7
Sapienic acid CH3(CH2)8CH=CH(CH2)4COOH cis-Δ6 16:1 n−10
Oleic acid CH3(CH2)7CH=CH(CH2)7COOH cis-Δ9 18:1 n−9
Elaidic acid CH3(CH2)7CH=CH(CH2)7COOH trans-Δ9 18:1 n−9
Vaccenic acid CH3(CH2)5CH=CH(CH2)9COOH trans-Δ11 18:1 n−7
Linoleic acid CH3(CH2)4CH=CHCH2CH=CH(CH2)7COOH cis,cis-Δ9,Δ12 18:2 n−6
CH3(CH2)4CH=CHCH2CH=CHCH2CH=CHCH2 cis,cis,cis,cis-
Arachidonic acid 20:4 n−6
CH=CH(CH2)3COOHNIST Δ5Δ8,Δ11,Δ14
Erucic acid CH3(CH2)7CH=CH(CH2)11COOH cis-Δ13 22:1 n−9
Triglycerides
As above stated, fats may be split in to glycerin and fatty acids, the resulting mixture containing three molecules of fatty
acid for each molecule of glycerin as shown in Fig 3.
Fig 3: Structure of Triglyceride

Since there are a number of different fatty acids that occur in natural fats, a great many different triglycerides are
encountered in nature. They include mono and diglycerides, free fatty acids, phosphatides, sterols, fatty alcohols, fat
soluble vitamins and other substances.
Groundnuts
The groundnut belongs to the pea and bean family and is a leguma but is considered as nut because of its high nutrition
value. The groundnut is the native of South America, where it has been cultivated for several countries. In 16th century
the Portuguese took it from Brazil to other countries and it spread in India also. Groundnut is the only nut that
grows beneath the earth. They become mature in about two months, when the leaves of the plant turn yellow. The
groundnut is particularly valued for its protein contents, which is of high biological value. Groundnut contains more
protein then meat, two and half more then eggs and more then any other vegetable food. The proteins in groundnut are well
balanced.
Groundnuts (Arachishypogaea), famous by its Indian name mongphaliis a species in the legume or "bean"
family (Fabaceae). The peanut was probably first domesticated and cultivated in the valleys of Paraguay.] It is an annual
herbaceous plant growing 30 to 50 cm (1.0 to 1.6 ft) tall. The leaves are opposite, pinnate with four leaflets (two opposite
pairs; no terminal leaflet); each leaflet is 1 to 7 cm (⅜ to 2¾ in) long and 1 to 3 cm (⅜ to 1 inch) across. The flowers are a
typical peaflower in shape, 2 to 4 cm (0.8 to 1.6 in) (¾ to 1½ in) across, yellow with reddish veining. The specific name,
hypogaea means "under the earth"; after pollination, the flower stalk elongates, causing it to bend until the ovary touches the
ground. Continued stalk growth then pushes the ovary underground where the mature fruit develops into alegume pod, the
peanut – a classical example of geocarpy. Pods are 3 to 7 cm (1.2 to 2.8 in) long, containing 1 to
4 seeds.
Fig 4: Groundnut Crop
Fig 5: Shelled groundnuts with skin
Groundnuts have all the benefits of a perfect fruit, in fact they can be considered in the same league as fruits when it comes
to contributing to our diet and health.
Health Benefit:
 Groundnuts contain five important nutrients namely food energy, protein, phosphorous, thiamin and niacin. It
maintains and repairs body tissues.
 Eating fresh roasted groundnuts with jaggery and goat's milk is very nutritious for growing children, pregnant women
and nursing mothers. It builds a resistance against all infections, such as Hepatitis and tuberculosis.
 Groundnuts contain 13 different vitamins (including Vitamin A, B, C and E) along with 26 essential trace minerals,
including calcium and iron.
 Groundnuts also contain zinc, good for protecting brain function, and boron, which helps to maintain strong bones.
 Ground nuts or groundnut products are useful in the treatment of hemophilia, and inherited blood diseases, which
cause hemorrhage. It is also useful in nose bleeding and in cases of excessive bleeding during menstruation in women.
 Groundnuts have good dietary fiber content so they are very good for digestion.
 Groundnuts are valuable in diabetes. It is also useful in diarrhea, especially chronic diarrhea, which is more frequent
immediately after a meal. The patient can use it by drinking goat's milk in which lemon is squeezed with a handful of
fresh roasted groundnuts.
Table 3: Nutritional Value of Groundnuts (per 100 grams)
Water 6.50gm
Energy 567kcal
Energy 2374Kj
Protein 25.80g
Fat 49.24g
Carbohydrate 16.13
Fibre 8.5 gm
Sugar, total 3.97gm
Calcium 93 mg
Iron 4.58mg
Magnesium 168mg
Phosphorus 376mg
Potassium 705mg
Sodium 18mg
Zinc 3.27mg
Copper 11.44mg
Manganese 1.934mg
Selenium 7.2 mcg
Groundnut Oil
Groundnut oil also known as peanut oil or arachisoil, is a mild tasting vegetable oil derived from groundnuts. The oil is
available in refined, unrefined, cold pressed, and roasted varieties, the latter with a strong peanut flavor and aroma,
analogous to sesame oil.
It is often used both for general cooking, and in the case of roasted oil, for added flavor. Groundnut oil has a high
smoke point relative to many other cooking oils, so is commonly used for frying foods. Its major component fatty
acids are oleic acid (46.8% as olein), linoleic acid (33.4% as linolein), and palmitic acid
(10.0% as palmitin). The oil also contains some stearic acid, arachidic acid, arachidonic acid, behenic acid,
lignoceric acid and other fatty acids.
Applications of Groundnut Oil:
o Peanut oil is high in monounsaturated “good” fat, and low in saturated “bad” fat, which is believed to help prevent heart
disease and lower cholesterol. It is also used to decrease appetite as an aid to weight loss. Some people use it to help
prevent cancer.
o Peanut oil is sometimes applied directly to the skin for arthritis and joint pain, dry skin, eczema, scalp
crusting and scaling without hair loss, and other skin disorders that cause scaling.
o Rectally, peanut oil is used in ointments and medicinal oils for treating constipation.
o Pharmaceutical companies use peanut oil in various products they prepare for internal and external use.
o In manufacturing, peanut oil is used in skin care products and baby care products.
Oil Extraction
The production process of vegetable oil involves the removal of oil from plant components, typically seeds. This can be
done via mechanical extraction using an oil mill or chemical extraction using a solvent. The extracted oil can then be
purified and, if required, refined or chemically altered.
Mechanical Extraction
Oils can also be removed via mechanical extraction, termed "crushing" or "pressing." This method is typically used to
produce the more traditional oils (e.g., olive, coconut etc.), and it is preferred by most "health-food" customers in the United
States and in Europe. There are several different types of mechanical extraction. Expeller- pressing extraction is
common, though the screw
press, ram press, and Ghani (powered mortar and pestle) are also used..
Solvent Extraction
The processing vegetable oil in commercial applications is commonly done by chemical extraction, using solvent extracts,
which produces higher yields and is quicker and less expensive. The most common solvent is petroleum- derived hexane.
This technique is used for most of the "newer" industrial oils such as soybean and corn oils. Groundnuts, Coconut, Palm,
Grape seed and Rice Bran are typically solvent extracted. Even the most perfect expellers leave at least six percent of oil in
the expeller cake. It is possible to recover these losses using a solvent extraction plant. Supercritical carbon dioxide can be
used as a non- toxic alternative to other solvents.
Enzymatic Pretreatment
The idea to develop an aqueous extraction process (AEP) to produce oil from oilseeds was born in the 1950s. In those
days, such a process seemed cheaper and less than the processes of extraction by solvent which supplied then the best
yields. Groundnut oil which contains 40~50% oil and 27-29% protein, is often used in cooking, because it has a mild flavor
and a relatively high smoke point. Due to its high monounsaturated content, it is considered healthier than saturated oils,
and is resistant to rancidity.The traditional approach for extracting oil from groundnut is solvent extraction by using
hexane. In the last few decades, aqueous (enzymatic) extraction has been attempted to extract oil/protein from many oil-
bearing materials, such as coconut, sun flower, rice bran, soybeans.The commercial hexane used as the most common
solvent for oil extraction is listed among hazardous air pollutants associated with neurological and respiratory disorders on
prolonged exposure (the International Standard Organization permits only 50 ppm residual hexane in oilseed meal).Hence,
there is a need to explore alternative safe and efficient oil extraction processes that may also result in edible protein.
II. MATERIALS AND METHODS
Groundnuts used in the experiments were purchased from the local market. Celluclast 1.5L was supplied by Novozymes
(Denmark). All the chemicals Hexane, citric acid and sodium citrate were of analytical grade and were procured from M/s.
Sd Fine chem. Pvt. Ltd., Mumbai.
2.1 Enzyme
Enzyme is a protein molecule acting as catalyst in enzyme reaction. Enzyme inhibition is a Science of enzyme- substrate
reaction influenced by the presence of any organic chemical or Inorganic metal or biosynthetic compound due to their
covalent or non-covalent interactions with enzyme active site.
The Enzyme used in the lab experiment is Celluclast1.5L. Celluclast is a lipase used for breaking down cellulose into
glucose, cellobiose and longer glucose polymers.
Fig 6: Purchased Celluclast 1.5L

2.1.1 Source:
Celluclast 1.5 L was manufactured by Novozymes was purchased.Cellulase that hydrolyzes (1,4)-beta-D- glucosidic
linkages in cellulose and other beta-D-glucans
2.1.2 Uses:
Celluclast can be used whenever the aim is to break down cellulosic materials into fermentable sugars, the reduction of
viscosity of soluble cellulosic substrates, or the increase in yield of valuable products of plant origin.
2.1.3 Activity:
Celluclast catalyzes the breakdown of the glucose polymers that comprise cellulose to glucose, cellobiose (i.e., pairs of
glucose units) and longer chains of glucose units. For practical purposes, the optimum conditions for activity of this enzyme
preparation are in the range pH 4.5- 6.0, and about 50-60 °C.
2.1.4 Storage:
Generally at a storage temperature of 25 °C, you can expect the enzyme to maintain its declared activity for at least 3
months. At lower temperatures (5-10 °C), the shelf life is considerably increased.
2.2 Hexane:
Hexane is significant constituents of gasoline. They are all colorless liquids at room temperature, odorless when pure, with
boiling points between 50 and 70 °C. They are widely used as they are easily evaporating non-polar solvents. Hexane is
used as solvent for the extraction of oil from oil seeds.
The "hex" prefix refers to its six carbons, while the "ane" ending indicates that its carbons are connected by single bonds.
Cooking oils are daily necessities used in all over the world and different types of oilseeds are grown at everywhere.
Besides food purpose, vegetable oil is also the source of bio-diesel, the new environmental friendly fuel. The cake from
oil press section could also be used as raw material for solvent extraction plant, the chemical way to extract oil. This kind of
extraction uses chemical solvent (Like Hexane) to dissolve oil content contained in cake or oil seeds. Oil is collected by
vaporizing solvent out which is later recycled. In configuring the solvent extraction plant, pre-pressing may be involved in
which case seeds are lightly pressed leaving about 14% to 18% oil in pressed cake. Solvent extraction will further process
these cakes and leave only 2% oil in the final cake (meal). This method results in higher capacity; Lower power
consumption, lower wear & tear / maintenance and high extract efficiency.
III. PHYSICAL CHEMICAL PROPERTIES OF
HEXANE
Physical state and appearance: Liquid.
Odor: Gasoline-like or petroleum-like (Slight.) Taste: Not available.
Molecular Weight: 86.18g/mole Colour: Clear Colourless.
pH (1% soln/water): Not applicable. Boiling Point: 68°C (154.4°F) Melting Point: -95°C (-139°F)
Specific Gravity: 0.66 (Water = 1) Vapour Pressure: 17.3 kPa (@ 20°C) Vapour Density: 2.97 (Air =
1)
Odor Threshold: 130 ppm
Water/Oil Distribution Coefficient: The product is more soluble in oil; log(oil/water) = 3.9
Iconicity (in Water): Not available.
Dispersion Properties: See solubility in water, diethyl ether, and acetone.
Solubility: Soluble in diethylether, acetone. Insoluble in cold water, hot water
2.3 Buffer Solution:
A buffer is an aqueous solution consisting of a mixture of a weak acid and its conjugate, or vice versa. Its
pH changes very little when a small amount of strong acid or base is added to it and thus it is used to prevent changes in
the pH of a solution. Buffer solutions are used as a means of keeping pH at a nearly constant value in a wide variety of
chemical applications.To prepare 0.1 M Sodium citrate 29.41 g was taken in 1liter Standard flask
and made up to the mark with distilled water and to prepare 0.1M citric acid 18.48 g was taken in another 1 liter Standard
flask and make up to the mark with distilled water.
From the above Standardized solutions, take 295 ml of 0.1 M sodium citrate and 20.5 ml of 0.1 M citric acid wastaken in
a1 liter standard flask and 1000 ml of distilled water was added up to the mark. The pH of the buffer was checked using
Thermo Scientific pH meter as shown below in Fig 7 and the pH was adjusted to 5 using 1N NaOH. If it is below the
desired pH add NaOH to raise it to the correct pH. If it is above the desired pH add phosphoric acid to lower it to the
desired pH [5.0]. Similar method was applied to prepare buffers with pH 5.5, 6.0.
Fig 7: pH meter
2.4 Experimental Setup:
The experimental set up consists of a hotplate showing a digital display of temperature and rpm. The maximum stirring
speed is 700rpm. The temperature can be set upto 180oC.
The ground nut powder is weighed and enzyme, pH buffer solution is added to a round bottom flask. A magnetic bead is
placed into the round bottom flask which helps in stirring. The temperature and RPM can be set as per our interest.
Fig 8: Experimental setup for study
2.5 Centrifuge:
Fig-9: Floor model Centrifuge

A centrifuge is a device for separating particles from a solution according to their size, shape, density, viscosity of the
medium and rotor speed, generally driven by an electric motor (or, in some older models, by hand), that puts an object in
rotation around a fixed axis, applying a force perpendicular to the axis.The centrifuge works using the sedimentation
principle, where the centripetal acceleration causes denser substances to separate out along the radial direction (the bottom
of the tube). By the same token lighter objects will tend to move to the top (of the tube; in the rotating picture, move to the
Center).
In a solution, particles whose density is higher than that of the solvent sink (sediment), and particles that are lighter than it
float to the top. The greater the difference in density, the faster they move. If there is no difference in density, the particles
stay steady. The maximum speed is 15,200 rpm and can hold up to 3L of samples.
2.6 Soxhlet extractor:
Fig 10: Soxhlet Extractor

A soxhlet extractor is a piece of laboratory apparatus invented in 1879 by Franz von soxhlet. It was originally designed
for the extraction of a lipid from a solid material. However, a soxhlet extractor is not limited to the extraction of lipids.
Typically, a soxhlet extraction is only required where the desired compound has a limited solubility in a solvent, and the
impurity is insoluble in that solvent. If the desired compound has a significant solubility in a solvent then a simple
filtration can be used to separate the compound from the insoluble substance. The sample is placed in the thimble.
Normally a solid
material containing some of the desired compound is placed inside a thimble made from thick filter paper, which is loaded
into the main chamber of the soxhlet extractor. The extraction solvent to be used is taken into a distillation flask and the
soxhlet extractor is now placed onto this flask. The soxhlet is then equipped with a condenser. The solvent is heated to
reflux. The solvent vapor travels up a distillation arm, and floods into the chamber housing the thimble of solid. The
condenser ensures that any solvent vapor cools, and drips back down into the chamber housing the solid material.
The chamber containing the solid material is slowly filled with warm solvent. Some of the desired compound will then
dissolve in the warm solvent. When the soxhlet chamber is almost full, the chamber is automatically emptied by a siphon
side arm, with the solvent running back down to the distillation flask. The thimble ensures that the rapid motion of the
solvent does not transport any solid material to the still pot. This cycle may be allowed to repeat many times, over hs or
days.During each cycle, a portion of the non-volatile compound dissolves in the solvent. After many cycles the desired
compound is concentrated in the distillation flask. The advantage of this system is that instead of many portions of warm
solvent being passed through the sample, just one batch of solvent is recycled. After extraction the solvent is removed,
typically by means of a rotary evaporator, yielding the
extracted compound. The non-soluble portion of the extracted solid remains in the thimble, and is usually discarded
4.7 Rotary Evaporator
Fig 11: Rotary Evaporator

A rotary evaporator is a device used in chemical laboratories for the efficient and gentle removal of solvents from samples
by evaporation.
Groundnuts
Enzymatic pretreatment
Centrifugati
on
Aqueous phase Solid phase

Oil phase
Drying
Rotary
evaporator
Soxhlet
Fig 12: Flowchart of the method.
Rotary evaporators are also used in molecular cooking for the preparation of distillates and extracts.
A simple rotary evaporator system was invented by Lyman
C. Craig.
1. The main components of a rotary evaporator are:
2. A motor unit that rotates the evaporation flask or vial containing the user's sample.
3. A vapor duct that is the axis for sample rotation, and is a vacuum-tight conduit for the vapor being drawn off of the
sample.
4. A vacuum system, to substantially reduce the pressure within the evaporator system.
5. A heated fluid bath (generally water) to heat the sample.
6. A condensate-collecting flask at the bottom of the condenser, to catch the distilling solvent after it re-condenses.
7. A mechanical or motorized mechanism to quickly lift the evaporation flask from the heating bath.
The vacuum system used with rotary evaporators can be as simple as a water aspirator with a trap immersed in a cold
bath (for non-toxic solvents), or as complex as a regulated mechanical vacuum pump with refrigerated trap. Glassware used
in the vapor stream and condenser can be simple or complex, depending upon the goals of the evaporation, and any
propensities the dissolved compounds might give to the mixture (e.g., to foam or "bump"). Commercial instruments are
available that include the basic features, and various traps are manufactured to insert between the evaporation flask and the
vapor duct. Modern equipment often adds features such as digital control of vacuum, digital display of temperature and
rotational speed, and vapor temperature sensing.
3.1 Enzymatic pre-treatment:
 The groundnuts were ground using a blender into fine granules.
 10 grams of blended groundnuts were weighed in each 250 ml round bottom flasks.
 To the above sample 7.5% of enzyme (i.e., 0.75g of enzyme) was dissolved in 50 ml of (Oil to buffer ratio of 1:5) pH
5.0 buffer solution was added.
 The seeds in parallel synthesizer was kept at 50oC for 5 hs at 450 rpm.
 The stirred round bottom flasks were then added 1.5ml of solvent (Commercial Hexane).
 And again were kept for continuous stirring for 15 minutes.
 Then the mixture in the round bottom flasks was transferred to centrifuge bottles.
 The round bottom flasks were given a hexane wash to avoid wastage of oil.
 Then the centrifuge bottles were kept in the centrifuge and speed was set to 8500rpm for 20 minutes.
 The centrifuge bottles were taken out after 20 minutes carefully without disturbing the layers.
 The oil and the cake were separated with the help of funnel and cotton.
 The Hexane layer was collected into the conical flasks.
 The cake was collected in petri dishes and was placed in the hot air oven bearing temperature around 110 oC.
 The cake was dried until it had no moisture in it.
 The Hexane layer and water were separated using separating funnels.
 The hexane which was evolved was kept rotor to evaporate the solvent and to extract oil.
 Then the cake after drying completely was scraped crushed into powder and was placed in thimbles.
 3-4 boiling chips were put into the round bottom flasks which were fitted with the soxhlet.
 The thimbles were then placed into the soxhlet and hexane was passed through the thimbles into the round bottom
flasks.
 Then the soxhlet apparatus was set and the temperature was set around 35oC.
 The soxhlet was run for about 4 h.
 Continuous water supply was given to condensers to avoid evaporation of solvent.
 After 8 h, soxhlet apparatus was turned off and was let to cool.
 Once it was cooled down, the solvent was taken and kept rotor to extract oil.
The received oil percentage was added to the percentage received in the hexane layer.
IV. RESULTS AND DISCUSSIONS
The effect of process parameters like concentration, temperature, pH, and reaction time on enzymatic pre- treatment was
studied. These parameters play a major role in the yield.
4.1 Effect of time
Reaction time plays a major role in the extraction of oil. The effect of time on enzymatic pre-treatment of
groundnuts were studied on varying time from 1-7 h and by keeping other parameters constant i.e., 7.5% of enzyme,
temperature at 50oC, 450 rpm of continuous stirring, oil to buffer ratio of 1:5 is shown Fig The free oil yield increased with
the increase in reaction time. After 1h of reaction 25.5% oil was extracted, by increasing time to 3h increased oil yield to
55.2%, increasing to 5h gave 73.1% oil yield and further increasing to 7 h 77.1% oil was extracted. As we can see in the
table that increase in reaction time was actually an increase in the oil
Table 4: Enzymatic Aqueous Oil Extraction by different reaction time
Reaction Time (h) Oil yield (%)
1 25.5
3 55.2
5 73.1
7 77.1
100
80
Oil yield (%)
60
40
20
0
0 2 4 6 8
Time
Fig 13: The reaction time was (h) by keeping other parameters constant. The oil yield increased with the increase in the
varied
reaction time.
4.2 Effect of enzyme
The effect of enzyme on enzymatic pre-treatment of groundnuts were studied on varying enzyme concentration from 1-10%
and by keeping other parameters at constant i.e., reaction time for 5hs, temperature at 50 oC, 450 rpm of continuous stirring,
oil and buffer in the ratio of 1:5. The oil yield increased with the increase in enzyme concentration.
The concentration of enzyme was varied to check the oil yield respectively. The experimental results showed that the oil
yield increased with the increase in concentration of
enzyme. The enzyme enhances the oil recovery from the groundnuts.
Table 5:Enzymatic Aqueous Oil Extraction by using different concentration of enzyme
Concentration of
Oil yield (%)
enzyme (%)
1 38.3
2.5 55.6
5 66.8
7.5 73.1
10 77.3
90
80
70
Oil yield (%) 60

50
40
30
20
10
0
0 2 4 6 8 10 12
Fig 14: The concentration of enzyme was varied by keep other parameters constants. The oil yieldof%enzyme
Concentration was more
(%)when the
enzyme concentration was more.
4.3 Effect of pH:
The effect of pH on enzymatic pre-treatment of groundnuts were studied on varying pH from 4.5-6.0 and by keeping other
parameters at constant i.e., enzyme concentration of 7.5%, reaction time for 5h, temperature at 50 oC, 450 rpm of
continuous stirring, oil and buffer in the ratio of 1:5. The activity range of pH for Celluclast is 4.5-6.0. pH range was varied
and the results were showed that oil yield was more at 5.0 pH.The below table shows the obtained oil% based on the pH
range.
Table 6:Enzymatic Aqueous Oil Extraction by varying pH buffer solutions.
pH Oil yield (%)
4.5 61.9
5.0 73.1
6.0 53
80
70
60
Oil yield (%)
50
40
30
20
10
0
0 2 4 6 8
pH
Fig 15: The aqueous oil extraction was done using 3 pH ranges i.e., 4.5, 5.0, 6.0. The results show that the enzyme activity
is more at pH 5.0 therefore resulting in higher oil liberation.
4.4 Effect of temperature:
The effect of temperature on enzymatic pre-treatment of groundnuts were studied on varying temperature from 50- 60 oC and
by keeping other parameters at constant i.e., pH 5.0,enzyme concentration of 7.5%, reaction time for 5hs, 450 rpm of
continuous stirring, oil and buffer in the ratio of 1:5.The activity range of temperature for Celluclast is 50-60 o C. The
experiment was performed for 3 different temperatures and the results were showing that the oil recovery decreased with the
increase in temperature. The below table shows the obtained oil% based on the temperature.
Table 7:Enzymatic Aqueous Oil Extraction by performing the experiment under different temperatures
Temperature(oC) Oil yield (%)

50 73.1
55 69.28
60 63.5
90
80
Oil yield (%)
70
60
50
40
30
20
10
0
0 20 40 60 80
Fig 16: Effect of varying temperatureo C)
Temperature( on oil recovery by aqueous enzymatic oil extraction. The oil % obtained from the
three different temperatures i.e., 50 o C, 55 o C, 60 o C
were recorded. The results signified that the oil recovery is more 50 o C as 73.1% oil was liberated.
V. CONCLUSION
Using Celluclast enzyme can increase the oil yield greatly compared to without enzyme. The optimal conditions for
enzyme should be favorable so as to enhance the pre- treatment. The time, concentration of enzyme play a major role in
pre-treatment as more the reaction time, more the oil yield will be and in the same way more the concentration of enzyme,
more the oil recovery will be from the groundnuts. The study confirmed that at pH 5.0 the oil yield is more when compared
to pH 4.5 and 6.0.The obtained results signified that the temperature which is more suitable for celluclast enzyme is 50 oC.
Enzymatic pre-treatment is one of the safest methods for oil extraction in both large scale and small scale. There are no
losses of nutrition and proteins in the oil extracted using enzymatic pre-treatment.
REFERENCES
[1] Dominguez H, Nunez M, Lema J. Enzymatic pretreatment to enhance oil extraction from fruits and oilseeds: a review. Food Chem
1994; 49: 271-86.
[2] Influence of enzymes on the oil extraction processes in aqueous media:Guillaume ricochon Lionel muniglia
[3] Aqueous Enzymatic Extraction of Oil and Protein Hydrolysates from Peanut: Yingyao Wang, Zhang Wang, Shangwei Cheng and
Fei Han 2008.
[4] Aqueous Enzymatic Extraction of Oil and Protein, Hydrolysates from Roasted Peanut Seeds,Shao Bing Zhang , Qi Yu
Lu ,Hongshun Yang, Yu Li, Shuai Wang 2010.
[5] Enzyme-Assisted Aqueous Extraction of Peanut Oil Aparna Sharma, S.K. Khare, and M.N. Gupta 2002.
[6] (Aqueous enzymatic extraction of peanut oil and protein hydrolysates Lihua Jiang, Di Hua, Zhang Wang, Shiying Xu, Food and
Bioproducts Processing, Volume 88, Issues 2–3, June–September 2010, Pages 233–238)
[7] (Response Surface analysis of enzyme assisted oil extraction factors for sesame, groundnut and sunflower seeds Singh R. K.; Sarker
B. C.; Kumbhar B. K. ; Agrawal Y. C. ; Kulshreshtha M. K.;Journal of food science and technology
,1999, vol. 36, no6, pp. 511-514)
[8] Gunstone, F.D., The Lipid Handbook, edited by F.D. Gunstone,J.L. Harwood, and F.B. Padley, Chapman and Hall, London,1986.
[9] Marlowe, I.T., T.J. Giddings, S.J. Richardson, and A. Stentiford,U.K. Industry and Ozone Pollution from Volatile Organic
CompoundEmissions, Warres Spring Laboratory– The Environmental Technology Executive Agency of the Department of Trade
and Industry, London, 1991, Report 878(PA).
[10] Hanmoungjai P, Pyle L, Niranjan K. Extraction of ricebran

oil using aqueous media. J Chem Tech Biot 2000;75: 348-
52.
[11] Van Eylen D, Indrawati M, Hendrickx M, Van Loey
A.Temperature and pressure stability of mustard seed
(Sinapisalba L.) myrosinase. Food Chem 2006; 97:263-71.
[12] Christensen FM. Enzyme technology versus
engineeringtechnology in the food industry.
BiotechnolApplBiochem 1989; 11: 249-65.

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INTERNATIONAL JOURNAL OF SCIENTIFIC PROGRESS AND RESEARCH (IJSPR) ISSN: 2349-4689

Issue 168, Volume 68, Number 01, February 2020

Extraction of Edible Oil From Groundnut By using

Fig 2: Structure of Cisand Trans-Isomers

Common name Chemical structure Δx C:D n−x

Myristoleic acid CH3(CH2)3CH=CH(CH2)7COOH cis-Δ9 14:1 n−5

Palmitoleic acid CH3(CH2)5CH=CH(CH2)7COOH cis-Δ9 16:1 n−7

Sapienic acid CH3(CH2)8CH=CH(CH2)4COOH cis-Δ6 16:1 n−10

Oleic acid CH3(CH2)7CH=CH(CH2)7COOH cis-Δ9 18:1 n−9

Elaidic acid CH3(CH2)7CH=CH(CH2)7COOH trans-Δ9 18:1 n−9

Vaccenic acid CH3(CH2)5CH=CH(CH2)9COOH trans-Δ11 18:1 n−7

Linoleic acid CH3(CH2)4CH=CHCH2CH=CH(CH2)7COOH cis,cis-Δ9,Δ12 18:2 n−6

Erucic acid CH3(CH2)7CH=CH(CH2)11COOH cis-Δ13 22:1 n−9

Fig 3: Structure of Triglyceride

Fig 4: Groundnut Crop

Fig 5: Shelled groundnuts with skin

Selenium 7.2 mcg

Fig 6: Purchased Celluclast 1.5L

Fig 8: Experimental setup for study

Fig-9: Floor model Centrifuge

Fig 10: Soxhlet Extractor

Fig 11: Rotary Evaporator

Aqueous phase Solid phase

Fig 12: Flowchart of the method.

Reaction Time (h) Oil yield (%)

Oil yield (%) 60

pH Oil yield (%)

Temperature(oC) Oil yield (%)

[10] Hanmoungjai P, Pyle L, Niranjan K. Extraction of ricebran

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