МИНИСТЕРСТВО ОБРАЗОВАНИЯ И НАУКИ РОССИЙСКОЙ ФЕДЕРАЦИИ
Федеральное государственное автономное образовательное учреждение
высшего образования
«Дальневосточный федеральный университет»
ШКОЛА БИОМЕДИЦИНЫ
Департамент пищевых наук и технологий
Дарвиш Фади Самир Юсра
Разработка способов стабилизации качества и сохраняемости растительных
масел с использованием натуральных антиоксидантов
МАГИСТЕРСКАЯ ДИССЕРТАЦИЯ
по образовательной программе подготовки магистров
по направлению 19.04.05 «Высокотехнологичные производства пищевых
продуктов функционального и специализированного назначения»
г. Владивосток
2018
ABSTRACT
The work contains 3 chapters, outlined on 91 pages, 27 tables, 25 figures, 102
bibliographic sources.
Keywords: sunflower oil, synthetic antioxidants, natural antioxidants, herbs and spices.
Purpose of work is the development of ways to stabilize the quality and the maintain
of vegetable oils using natural antioxidants.
The results of research proved the usefulness of using herbal extracts as natural
antioxidants to reduce the process of oxidation of vegetable oils and used as substitutes
for industrial antioxidants. After making extracts of different herbs (thyme, ginger,
rosemary,sage) using different solvents(ethanol 95%, ethanol 70%, propylene glycol
95%, ethanol 95%, and propylene glycol 95%) we measure the total amount of phenolic
compounds and the free radical scavenging activity (DPPH) of herbs extract. These
extracts were either added separately or added a mixture of these extracts to refine
sunflower oil and after measuring the value of peroxide and the acid values of the different
oil samples every 15 days for two months and comparing with the acid and peroxide value
of either the sample of sunflower oil without additions or the sample of sunflower oil with
BHT. The results of experiments show that all the extracts have an antioxidant effect
higher than the industrial antioxidant. In addition, the sample containing the rosemary and
thyme extract had the lowest peroxide and acid value by comparing it with other oil
samples, and this shows that this mixture of extracts has the ability in delaying the
occurrence of the oxidation process of refined sunflower oil. Therefore, we suggest that
the rosemary and thyme extract can be used as a potential natural antioxidant for vegetable
oils rather than industrial antioxidants
1
CONTENT
INTRODUCTION ............................................................................................................ 4
CHAPTER 1 LITERATURE REVIEW ...................................................................... 5
1.1 Lipids oxidation ........................................................................................................ 5
1.2 Antioxidants .............................................................................................................. 7
1.2.1 Antioxidants found in Edible oils ........................................................................ 10
1.2.2 Synthetic Antioxidants ......................................................................................... 12
1.3 Natural antioxidants ................................................................................................ 15
1.3.1 Spices and herbs ................................................................................................... 15
1.3.2 Natural antioxidants properties ............................................................................ 19
1.4 Oxidative stabilization of vegetable oil by adding natural antioxidants ................ 20
1.4.1 Using the rosemary extract as a natural antioxidant for vegetable oil ................. 24
1.4.2 Using the sage extract as a natural antioxidant for vegetable oil ........................ 26
1.4.3 Using the thyme extract as a natural antioxidant for vegetable oil ..................... 28
1.4.4 Using the ginger extract as a natural antioxidant for vegetable oil ..................... 29
1.4.5 Using the oregano extract as a natural antioxidant for vegetable oil ................... 31
CHAPTER 2 OBJECTS AND METHOD OF RESEARCH ............................... 33
2.1 Research objectives ................................................................................................. 33
2.2 The preparation of herbs extracts using ethanol 95% ............................................. 34
2.3 The preparation of herbs extracts using ethanol 70% ............................................. 34
2.4 The preparation of herbs extracts using propylene glycol 95% ............................. 35
2.5 The preparation of herbs extracts using propylene glycol 95% and ethanol 95% . 36
2.6 Determination of the content of polyphenols for herbs extracts............................. 37
2.7 Measuring the free radical scavenging activity (DPPH) of herbal extracts ........... 38
2
2.8 Determine the peroxide value of oil samples .......................................................... 39
2.9 Determine the acid value of oil samples ................................................................. 40
CHAPTER 3 RESULTSAND DISCUSSION .......................................................... 41
3.1 The amount of phenolic compounds for herbs extracts .......................................... 41
3.2 The DPPH radical scavenging ability of the herbs extracts ................................... 42
3.3 The peroxide values and acid values of refined sunflower oil samples ................. 43
3.3.1
Effect of adding herb extracts extracted with ethanol 95% separately or BHT on
the peroxide values and on acid values of refined sunflower oil samples .................... 43
3.3.2
Effect of adding a mixture of herb extracts extracted with ethanol 95% or BHT on
the peroxide values and on acid values of refined sunflower oil samples .................... 47
3.3.3 Effect of adding herb extracts extracted with ethanol 70% separately or BHT on
the peroxide values and on acid values of refined sunflower oil samples .................... 52
3.3.4 Effect of adding a mixture of herb extracts extracted with ethanol 70% or BHT
on the peroxide values and on acid values of refined sunflower oil samples ............... 56
3.3.5 Effect of adding herb extracts extracted with ethanol 95%and propylene glycol
95% or BHT on the peroxide and on acid values of refined sunflower oil samples .... 61
3.3.6 Effect of adding a mixture of herb extracts extracted with ethanol 95%and
propylene glycol 95% or BHT on the peroxide values and on acid values of refined
sunflower oil samples.................................................................................................... 65
3.3.7 Effect of adding herb extracts extract with propylene glycol 95% separately or
BHT on the peroxide values and on acid values of refined sunflower oil samples ...... 70
3.3.8 Effect of adding a mixture of herb extracts extract with propylene glycol 95% or
BHT on the peroxide values and on acid values of refined sunflower oil samples ...... 74
CONCLUSIONS............................................................................................................. 79
LIST OF SOURCES USED........................................................................................... 81
3
INTRODUCTION
Relevance of the topic. The oxidation of vegetable oils is considered one problem
in the food industry, thus, are added antioxidants in food [1].
The oils and fats are exposed to the phenomenon of rancidity which lead to a decline
in value and quality deterioration oils and lipid containing foods, especially when exposed
to elevated temperature, light and oxygen [2].
Long time ago synthetic anti-oxidants such as butylated hydroxyanisole (BHA) and
(BHT) were used, to lengthen the period of food oils conservation but it was found that
these antioxidants have negative effects on the health of the consumer[3].
So there is a need to look for antioxidants from natural sources as an alternative to
prevent the degradation of fats and oils and safe for the consumer.
There are many of the plant extracts containing natural antioxidants, such as, ginger,
spices, herbs, seeds, rosemary, cereals, cocoa shell, grains, fruits, catnip, hyssop, oregano,
sage and thyme and other natural sources[4].
Where the interest in the use of herbs and spices has increased as a source of natural
antioxidants because they contain many phytochemical compounds such as phenolic
diterpenes, flavonoids, alkali, tannins and phenolic acids [5].
From the consumer's perspective, these antioxidants are safe in terms of health and
did not need to safety followed tests as synthetic anti-oxidation because they are consumed
from natural sources long ago [2, 6].
Purpose of work–Development of ways to stabilize the quality and the maintain of
vegetable oils using natural antioxidants.
4
CHAPTER 1 LITERATURE REVIEW
1.1 Lipids oxidation
The oxidation of lipids affects not only the nutritional value and life of food
products containing oils, destruction of contained vitamins (the fat-soluble A, D and E),
but may also affects human health [7].
Oxidative rancidity takes place in oils when these oils are exposed to elevated
temperature, oxygen and light or other catalysts cause unsaturated fatty acids to turn into
free radicals. This happens when hydrogen is lost from the a-methylenic carbon in the
fatty acid group [8].
Oxidation reactions lead to the production of free radicals which may in turn begin
chain reactions that give rise to more oxidation by chain reaction [9].
These free radicals are easily oxidized to produce hydroperoxides and organic
compounds, which are responsible for the obnoxious odors and flavors characteristic of
rancid fats in vegetable oils [10].
The oxidation of lipids which takes place in the triacylglycerol molecules includes
3steps [11].
The first phase of fat oxidation is called initiation .This phase is called the phase of
the production of free radicals fatty acids. This process occurs when the hydrogein atom
is removed from unsaturated fatty acids specifically from the group of methylene alkylic.
This reaction is often caused by the presence of light, transition metals or high temperature
[12].
The interactions and changes that occur at this stage are explained in the following
equations (1, 2).
The second phase of fat oxidation is called propagation .The spread of fat free
radical in oxidation processes occurs by a chain reaction that consumes oxygen and
5
produces new radical types such as peroxyl radicals, ROO or by forming peroxides,
ROOH .Products R • and ROO• can increase the propagation of free radical reaction and
ROO• Initiate chain reaction with other molecules creating hydropyroxides fat and free
fat radical.
This reaction is repeated several times so that it produces and accumulates large
amounts of heproperoxides and stops only when the unsaturated fat or fatty acid molecules
are depleted. The interactions and changes that occur at this stage are explained in the
following equations (3, 4,5,6,7,8).
The final phase of fat oxidation is called termination .Radical interaction ends when
radicals react and produce a non-radical product. This happens because radicals are very
reactive and when there is a decrease in the amount of fatty acids, the bonds are radical to
each other and form a non-stable stable compound. This will terminate the interaction
[13].
The interactions and changes that occur at this stage are explained in the following
equations (9, 10, 11).
(1) RH →R•+H•
(2) ROOH+ RH → R•+H2O+RO•
(3) R• + O2 →ROO•
(4) ROO• + RH →ROOH + R•
(5) ROO• + R → ROOR•
(6) ROOR•→RO•+RO•
(7) RO• +RH→ROH +R•
(8) RO• +O2→R"O+HO2•
(9) ROO• +ROH → ROOH + R"O +HO•
(9) R• + R• →R-R
(10) ROO•+R•→ROOR
(11) ROO• + ROO• →ROH + R"COR + O2
6
The mainly oxidation compound are hydroperoxides [14]. These compounds
quickly are transformed into other compounds because they are unstable such as
aldehydes alkanes, alcohols, and acids which called secondary products [9].
There are other ways to explain the mechanism of fat oxidation:
The first method is called autoxidation. Autoxidation is the most common reaction
to explain the mechanism of fat oxidation. This reaction occurs through the incorporation
of unsaturated fatty acids with molecular oxygen to produce hydroperoxides.
The other way occurs in the presence of oxygen, when unsaturated fats are exposed
to light and a sensitizer this leads to the production of alkyl hydroperoxide. This
interaction is called photooxidation[15].
1.2 Antioxidants
There are several ways to inhibit the oxidation process such as denial of access to
oxygen, use of lower temperature, inhibition of enzymes catalyzing oxidation reduce the
oxygen pressure and the use of suitable packaging [16].
Another way to protect against oxidation use specific additives called antioxidants
[17].
Antioxidants are substances obtained from natural sources or industrial sources
added in small quantities to vegetable oils to improve oxidative stability, reduce the
oxidation rate or delaying the occurrence of auto-oxidation process by preventing free
radicals formation [9, 8,18].
Antioxidants lengthen the induction period of oxidation, or slow the rate of
oxidation. Antioxidants disable free radicals, such as alkyl or peroxyl fat radicals, inhibit
the effect of transitional metals, put out the shirt, oxygen and disrupt the sensitizers [19].
Antioxidant additives must be chosen to edible oils for the purpose of keeping
unsaturated fatty acids to get better stability to thermal degradation which takes place
between 150 and 220 °C [20, 21].
7
Antioxidants can give a hydrogen atom to free radicals and turn it into a more stable
non-radical product. The main contributors to antioxidants and hydrogen are mono or
polyhydroxenolic compounds with different substitutes in the aromatic nucleus. The
standard recovery capacity of one of the alkyl, peroxyl and alkoxy radicals of PUFA is
600, 1000, 1600 mph, respectively. Usually the usual recovery of antioxidants is 500 mph
or less.
This clearly indicates that antioxidants interact with radical peroxyl prior to a
radical pyroxyl reaction with another fatty molecule leading to the formation of other free
radicals. Any radical of antioxidants formed by interaction with the peroxyl radical of fat
has less energy than radical peroxyl.
Antioxidants are divided into two types depending on its source natural antioxidants
and industrial antioxidants [22].
Because of high performance and low cost for synthetic antioxidants, they are
widely used in food oil to reduce rancidification such as butylated hydroxyanisole
(BHA),octalgallate(OG),
butylated
hydroxytoluene
(BHT)
and
2,4,5-
trihydroxybutyrophenone (THBP) , but it was found that these antioxidants have negative
effects on the health of the consumer [9,23].
There are many plants which have antioxidant properties such as ginger, spices,
herbs, teas, rosemary, cereals, cocoa shell, grains, fruits, catnip, hyssop, oregano, sage and
thyme and other. Antioxidant effects are due mainly to contain these plants on phenolic
compounds [24, 25].
Antioxidants depending on the mechanism of action are divided into primary and
secondary antioxidant [26, 27].
In order to get a synergistic effect both types can be mixed with each other.
However some antioxidants have more than one way of action [28].
So it was important to use antioxidants that have the ability to delay and protect the
oil from the emergence of rancidity in fatty foods; they are working to remove the active
8
oxygen forms, which are considered the first step oxidize or demolition oxidative, through
interact the free radical of fatty acids with antioxidants mutant to stable form.
These antioxidants should be safe to use, and have no odor or flavor, and added
in very small quantities, low costs [14].
In the initiation stage the antioxidants function prevent the formation of free
radicals. However in the propagation stage antioxidants function by donating hydrogen to
terminate the free radical chain [10].
The primary antioxidants act as follows:
-It can end the free radicals chain through donation hydrogen to free radicals and
turn them into stable compounds.
-By reacting with lipid radicals they are able to form lipid-antioxidant complexes.
-By reacting with a lipid free radical they are able to inhibit the initiation step.
-By reacting with proxy they are able to delay propagation step [9].
The Secondary antioxidants act as follows:
-Convert hydroperoxide to non-radical type.
-Disrupt singlet oxygen.
-Oxygen scavengers.
-By absorbing ultraviolet radiation.
-By giving primary antioxidants H+[9].
Table 1 – Different classes of antioxidants [29]
Class of antioxidants
Examples
Function
Free radical scavengers
BHA
Block free radicals by
BHT
donating a hydrogen atom
TBHQ
Propyl gallate tocopherols
Extracts from spices and
herbs (rosemary, clove,
sage, oregano)
Oxygen scavenger
Ascorbic acid
React with oxygen
9
Erythorbic acid
Ascorbate
Sulfites, bisulfites
Ascorbic palmitate
Chelating agents
Citric acid
React with metal ions
EDTA
capable of catalyzing
Phosphates
oxidation
1.2.1 Antioxidants found in Edible oils
Some antioxidants naturally exist in edible oils such as phenolic compounds
tocopherols, sterols, carotenoids, and tocotrienols.
Tocopherols were considered the mainly antioxidants exist in edible oils. Some
vegetable oils, such as sunflower, soybean, corn oils, and canola contain high
concentrations of tocopherols [30].
During the process of removing odor from the oils part of the tocopherols removed,
the remaining amount of tocopherol remains sufficient to protect against oxidation.αtocopherol comparison of the types of tocopherol it is considered more active antioxidant
[31, 32].
Other antioxidants present in some oil belong to group of lignans which contain
phenolic acids. During processing of oil seeds many of them are insoluble.
During processing of oil seeds, natural antioxidants are found in oil seeds partition
into hydrophilic fractions or liposoluble.
Through the solvent extraction or expeller pressing many of lipophilic antioxidants
are transported into crude oil by extraction [31].
Many previous research has indicated that oil peroxy radicals are able to interact
with tocopherols faster (104–109M-1.S-1) this is compared to their interaction with lipids
(10–60M-1. S-1), so they are able to chain breakers and radicals scavengers [33].
10
103 to 108 polyunsaturated fatty acid can be protected by1 tocopherol molecule at
low pv (peroxide value).H atom at the 6-OH on tocopherols ring can transfer to oil peroxy
radical subsequently scavenge this peroxy radicals [30].
Effectiveness of tocopherols as antioxidants related to its concentration and isomers
[30].
Phenolic antioxidants through interaction with ROO• can produce ROOH and
phenoxyl radical which is relatively stable.
ROO· +PhH →
ROOH + Ph·
Ph· through interaction with ROO• can produce non-radical products
ROO·+
Where
Ph· → Non radical products
ROO•: peroxyl radicals
Ph·: phenolic antioxidants [33].
Carelli and others found that when a- tocopherol added to sunflower oil for the
purpose of increasing oxidative stability. When the amount of tocopherol is increased, the
oxidative stability increased. Compared with the industrial antioxidant (BHT) show that
its effectiveness similar to the effectiveness of industrial anti-oxidant in sunflower oil [34].
Table 2 – Main classes, compounds and mechanisms of action of antioxidants
present in edible vegetable oils [35]
Classes of antioxidants
and selected compounds
Tocols
Antioxidant mechanism
Primary or chain breaking antioxidants
α-tocopherol
c-tocopherol
Phenolic compounds
Primary or chain breaking antioxidants;
Hydroxytyrosol (α)
Also secondary or preventive antioxidants2
Tyrosol
acting as chelators of metal ions;
Might stabilize and prevent decompositionof
hydroperoxides.
11
Carotenoids
Secondary or preventive antioxidants
β-carotene
acting as singlet oxygen quenchers;
Also primary or chain breaking antioxidants.
Phytosterols
β -sitosterol
Possible primary or chain breaking
antioxidant
1.2.2 Synthetic Antioxidants
In many countries, industrial antioxidants are used, but there are many uncertainties
about their health effects.
The main reasons in adding these substances to food oils are because of the ability
of industrial antioxidant to prolong the life of food oils by delaying or inhibiting the
process of oxidation [36].
BHT and BHA these antioxidants can be added alone to food or with other
chemicals that have the properties of antioxidants such as ascorbic acid, phosphoric acid,
propyl gallate and citric acid.
(Butylated
hydroxyanisole;
C11H16O2)
and
(butylated
hydroxytoluene;
C15H24O), as mentioned earlier, are commonly used preservatives in foods with fats and
oils [38]. Their purpose is mainly to delay the oxidation of foods so that they do not change
color, flavor, or odor over time. They do this by reacting with oxygen before they react
with fats and oils so that fats and oils do not oxidize and spoil.
Long time ago compounds such as TBHQ, BHT and BHA have been known to be
antioxidants from unnatural sources, they were produced in laboratories [37, 38].
BHA the scientific name is butylated hydroxyanisole is one of the most widely used
antioxidants due to its high solubility in fats and oils and is able to stabilize at high
temperatures. And it is added to many foods including vegetable oil, frying oil.
12
The use of BHA as an antioxidant increased because it has the ability to stabilize
at high temperatures. Even though it is used less than BHT [33] .
Figure 1 - Chemical Composition of BHA
BHT the scientific name is butylated hydroxytoluene. Although all the benefits of
BHT and its derivatives, this family of interesting aromatic compounds very well known
as promising antioxidants.
There are more than publications are discussed exclusively on the properties of
antioxidants Of BHT and its derivatives.
Currently, BHT is one of the antioxidants widely used in the food industry.
It is used in low-fat food, fish products. It is also widely used in combination with
other antioxidants such as, propyl gels, and citric acid to stabilize of oils and high-fat
foods.
13
Figure 2 - Chemical Composition of BHT
The action of this industrial antioxidant is similar to BHA as it works to reduce
oxygen radicals and stop the spread of oxidation operations. But its use in food oil is lower
compared to BHA.
Because it is unstable at high temperatures this makes it suitable for use in food
oils at moderate temperatures [33].
TBHQ the scientific name is tert-butylhydroquinone. TBHQ this compound can be
especially used as an antioxidant for oils with high content of unsaturated fatty acids and
can be added alone or added with other antioxidants such as BHA or BHT.
It possesses some advantages compared with other antioxidants in prolonging the
storage period of edible oils [37].
14
Figure 3 - Chemical Composition of TBHQ
Propyl gallate this type can get from natural gallic acid .It has high antioxidant
activity in vegetable oils. Compared with BHT and BHA which has low solubility in oils.
Antioxidant activity of (PG) belongs to its content of trihydroxy substitutions and
the para-OH group and ester group is considered the most important one.
Similar to other antioxidants gallate esters prevent fat peroxidation through the
transfer (h) atom from a phenolic (oh) group to lipid radicals.
During this process relatively stable compounds will be formed due to the ability
of free radicals of (pg) to correlate with lipid radicals. For this reason this compound can
be considered effective antioxidant [39].
1.3 Natural antioxidants
1.3.1 Spices and herbs
Spices and herbs are added to food in order to give them flavor [40].Interest has
increased in the use of these substances in the food industry because of their ability to
delay oxidative degradation of oil and increase nutritional and life of food because of their
15
antioxidant properties which are superior to many of the natural anti-oxidants and
synthetic antioxidants currently used [41].
Because of high contents of phenolic compounds, they have a high ability to donate
hydrogen atoms [40].
These advantages are due to many compounds, including, carotenoid sterpenoids,
vitamins, flavonoids, minerals, phytoestrogens which make spices and herbs used for food
preservation [16].
There are natural antioxidants that are added to foods such as tocopherols, citric
acid, tocotrienols, ascorbic acid, and enzymatic antioxidants.
Nevertheless these natural antioxidants have some flaws, synthetic antioxidants
higher antioxidant activity as compared with natural antioxidants.
Although they have advantages such as consumers readily accept them and
consider to be safe, natural antioxidants are accepted and currently available by health
experts [42].
Herbs and spices such as, oregano, clove, savory, marjoram, sage, garlic, thyme,
cinnamon, basil, nutmeg, pepper, turmeric, cumin and rosemary are considered a good
source of antioxidants.
Table 3 – Antioxidants isolated from herbs and spice [43, 44]
Spice/herb
Scientific
Antioxidant compounds
name
Rosemary
Mode of
action
Rosemarinus
Carnosol, carnosic acid,
Scavenge
officinalis
rosmanol,
superoxide
rosmadial, diterpenes
radicals, lipid
(epirosmanol,
antioxidant,
isorosmanol,
and metal
rosmaridiphenol),
chelator
rosmariquinone,
rosmarinic acid
16
Sage
Salvia
Carnosol, carnosic acid,
Free radical
officinalis L.
rosmanol,
scavenger
rosmadial, methyl and
ethyl esters
ofcarnosol,rosmarinic
acid
Oregano
Origanum
Rosmarinic acid, caffeic
Free radical
vulgaris
acid,
scavenger
protocatechuic acid, 2caffeoyloxy3-[2-(4-ydroxybenzyl)4,5dihydroxy]
phenylpropionic acid;
flavonoids—apigen,
eriodictyol,
dihydroquercetin,
dihydrokaempherol;
cavacrol, thymol
Thyme
Thymus
Thymol, cavacrol, p-
Free radical
vulgaris L.
Cumene-2,3-
scavenger
diol, phenolic acids
(gallic acid,
caffeic acid, rosmarinic
acid),
phenolic diterpenes,
flavonoids
Gingerol,shogaol,
zingerone
17
Active components are found in herbs and spices such as phenolic diterpenes and
mono (carnosol,carnosic acid, thymol rosmadial, cavacrol, and rosmanol,) derivatives and
phenolic acids such as caffeic, gallic, protocatechuic, ferulic, and rosemarinic.
Components belong to gingerol such as shagoal and gingerol, diarylheptanoids
cassamunin A, B, C and curcumin, phenolic amides compounds such as capsaicinol and
capsaicin and flavonoids components (isoharmnetin , kaempferol ,luteolin, apigenin, , and
quercetin) [45].
Figure 4-Antioxidative phenolic found in plants
Herbs which belong to lamiaceae family contain a compound called rosmarinic acid
.This composite is capable of scavenging (DPPH potential) that's because it contains 4OH. This compound is present at a high concentration in oregano about 500ppm, 30000
ppm in peppermint and 37000 ppm in lemon balm [33].
18
Figure 5 -flavonoids components found in plants [95]
1.3.2 Natural antioxidants properties
Natural antioxidants must have the following properties:
-Must be safe to eat by the consumer.
- Should not leave harmful effects after ingestion.
- Should not leave any odor or taste unacceptable in the oils that are added to it.
- They should be effective when used with low concentrations.
- They must maintain its stability during the processing and conservation of the oils
to which it is added.
- They must be cheaply priced.
- They should be easy to use and easy to handle.
19
- They must be able to dissolve in the oils that are added to it.
- They should be able to prolong the life of the oils that are added to it [43].
1.4 Oxidative stabilization of vegetable oil by adding natural antioxidants
Allam and others conducted experiments to measure the thermal stability of
sunflower oil heated to 180 degrees celsius for an hour after adding a combination of
natural antioxidants (mixed mono- acylglycerol citrate and tocopherols) and industrial
antioxidants such as (TBHQ, AP, BHA, PG, and BHT) the results showed that the oil
containing TBHQ has the highest thermal stability while the least is the oil containing AP
[46].
Several previous studies have indicated that the peroxide value of sunflower oil
containing rosemary extract and stored for 8 hours at 98 0С(16meq O2/kg of oil) is less
than twice the value of peroxide of sunflower oil without adding (32meq O2/kg of oil).
Babović and others found that the effectiveness of the industrial antioxidant (BHA)
and sage extract after adding each of them to the heated sunflower oil for temperature 98
for 12 hours have the same effectiveness [47].
Saba Ajeena and others found that after measuring radical scavenging of the
extracts of each, cumin fruits, black pepper seeds, and sage leaves and Industrial
antioxidants (BHT and BHA) the results showed that the sage extract has the best ratio
(49.97%) then fruits of cumin 47.88%, while BHA 36.77 and BHT 46.97% while the
lowest ratio was black pepper seeds 2.95% [48].
When adding the extract of nigella seeds to sunflower oil at temperatures 60 and
100 and room temperature, this extract showed oxidation activity, which may be due to
containing phenol compounds. Also adding this extract in 1000 ppm has been shown to
be most effective as an antioxidant [49].
20
Khalil Dhouib and others found that the effectiveness of the industrial antioxidant
(BHT) at 200 ppm and basil extract at 300-400 ppm after adding each of them to the
sunflower oil have the same effectiveness [50].
Marinova and others concluded that the addition of myricetin extract to sunflower
oil at temperature 100 showed activity as an antioxidant better than α-tocopherol [15].
Some previous studies have indicated that the extract of garlic extracted by
methanol when added to sunflower oil at concentrations 500 and 1000 ppm and compared
to industrial antioxidants (BHA and BHT) at 200 ppm that were added to the same oil.
The extract with concentrations 1000 ppm has been shown more effective as an
antioxidant than industrial antioxidants [51].
Imran and others found through the experiments that the addition of extracts of
Fenugreek, Liquorice and Mint to sunflower oil stored at room temperature to increase
the stability of this oil have shown almost equal effectiveness of industrial antioxidants
(BHA and BHT) [52].
Mostafa Taghvaei and others found that the addition of extracts of olive cake with
concentrations 200ppm to sunflower oil stored at room temperature in order to increase
the stability of this oil against oxidation and prolong the life of this oil has shown more
effective than BHT [51].
Several studies have indicated that when adding the extract of sesame cake to
sunflower oil. This extract has an antioxidant effect higher than industrial antioxidants
[51, 53, 54].
Sameera and others found that when adding the extracts of oleoresin curcuminoids
, lecithin and capsicum to sunflower oil ,these extracts have been shown to be effective as
an antioxidant similar to the industrial antioxidant TBHQ which was added to the same
oil with a concentration200ppm [55].
Sesamin, sesamol, and sesaminol are considered as lignan compounds present in
sesame oil. Many previous studies have indicated that comparison between effectiveness
of sesamol and sesaminol as an antioxidant in sunflower oil and effectiveness of sesamin
21
found that the latter is less effective in autoxidation of sunflower oil and it is also less at
scavenge radicals. Not only in the light but also in the dark, sesamol plays the role of antioxidation during the oxidation process [40].
Some studies have indicated that carnosic acid, camphor, rosmarinic acid and
carnosol, are responsible for the efficacy of sage extracts as antioxidants [56].
Some studies have indicated that several antioxidant compounds can be obtained
from sage such as luteolin-7-O- β -glucopyranoside, 6-O-caffeoyl- β -D-fructofuranosyl(2→1)- α -D-glucopyranoside, 1-O-caffeoyl- β -D-apiofurano-syl-(1→6)- β -Dglucopyranoside, and 9-ethylrosmanol ether. But the more active compounds are
rosmarinic acid carnosic acid and carnosol [57].
Tea is an evergreen tree belong to the theaceae family, exist in several countries
[58, 21].
Polyphenols of tea at a concentration 200 mg/kg have been added to salad oil, then
they were stored for45 days have shown efficacy in preventing oil degradation, but there
was a change in color [43].
Green tea has antioxidant effectiveness as a result of containing compounds such
as flavonoids, vitamins, and tannins. This activity is mainly due to the phenol compounds
it contains which is estimated at 450 mg/g as a catechins which consists mainly of
derivatives of gallic acid [40].
Taghvaei and others have found through experiments that when added green tea
extract with concentration100, 200, 500 and 1,000 ppm to each of the menhaden and seal
blubber oil and in comparison with the addition of both BHA, TBHQ and BHT with 200
ppm and α-tocopherol at 500 ppm, the efficacy of green tea extract at concentration
200ppm as an antioxidant is better than α-tocopherol, BHT and BHA but less than TBHQ
[59].
Chen and others have found through experiments that when added green tea extract
at a concentration of 0.02% to rapeseed oil and in comparison with the addition of both
BHT and rosemary extract.
22
They found that the efficacy of green tea extract at concentration 0.02% as an
antioxidant is better than BHT and rosemary extract.
Anna Gramza and others found that extract of green tea extracted by ethanol and
when added to sunflower oil with concentration1000ppm show antioxidant activity better
than α-tocopherol [58].
Basil this type of spices belongs to a family Lamiaceae. It is usually used for
cooking or for medical purposes. There are many compounds that are highly active as
antioxidants found in high concentrations in basil extract such as phenolics and eugenol.
It is also found some phenolic acids such as caffeic, vanillic, p-coumaric, syringicand
ferulic acid [60].
KritiSoni and Kanchan Kohli found that rosmarinic acid is the main phenolic
compound found in basil leaves and was believed to be responsible for the effectiveness
of basil as an antioxidant.
It was also found that essential oil of basil contains some compounds such as, epiα-cadinol, γ –cadinene, α-bergamotene and, linalool [61].
Khalil Dhouib and others through experiments found that the basil extract has the
ability to increase the stability of sunflower oil when added to it because it has the ability
to improve hydrolytic stability and prevent the double bond conjugation.
When added basil extract at concentrations 200–500 ppm to sunflower oil, the
results showed that has the ability to maintain the stability of this oil more than industrial
antioxidants such as BHT [62].
Some previous studies have indicated that when added basil extract to soybean oil,
it has the ability to protect this oil from oxidation more than industrial antioxidants such
as TBHQ [60].
Svitlana and others found that when added basil extract to sunflower oil, when
increase the amount of added extract from 2 to 10 % this led to increased antioxidant
activity by 1.57 times [63].
23
1.4.1 Using the rosemary extract as a natural antioxidant for vegetable oil
This type of herb belongs to the Lamiaceae family .Rosemary was first planted in
the vicinity of the Mediterranean Sea but today it is cultivated in many parts of the world
as an ornamental and aromatic plant.
Leaves of this plant It is usually added to foods to improve the taste, but this plant
has also been widely used for various medicinal purposes in traditional medicine [64].
The leaves of rosemary used as ingredient for giving flavor for food
productsandbecause of health useful properties such as antirheumatic, antialgesic and
antimicrobia effects has been known as beneficial plant for the health.
Studies have indicated that antioxidant activity of α-tocopherol is less than
rosemary extract [65].
Because of components such as rosmaridiphenol, rosmanol, carnosoand
rosmariquinone,which are present in rosemary oleoresin the effectiveness of anti-oxidants
BHA are less four times than rosemary as antioxidant [66].
Carnosic acid this component, according to several previous studies, indicates that
it is more active as antioxidant compared to other component found in rosemary extract
[72, 67].
Some previous studies have indicated that the extract of rosemary when added to
vegetable oil at concentrations 400 ppm and compared to industrial antioxidants (BHA
and BHT) at 200 ppm that were added to the same oil.
The extract at concentrations 400 ppm has been shown more effective as an
antioxidant more than industrial antioxidants [67].
Because of higher concentration of phenolic compounds antioxidant activity of
rosemary isattributed to contain the phenolic compounds such as osmanolcarnosic acid,
isorosmanoepirosmanol and carnosol.
24
Figure 6 -Some anti-oxidant compounds are present in rosemary [68]
BHA and BHT known as synthetic antioxidants are able to donate hydrogen found
in a group oh in a single aromatic ring. However carnosic acid contains a single aromatic
ring with two groups OH and that are able to donate H [40].
Xiaoqiang Chen and Ying Zhang found through experiments that the extract of
rosemary when added to sunflower oil at concentrations 200 ppm and compared to
industrial antioxidants (BHA and BHT) at 200 ppm that were added to the same oil.
25
The extract at concentrations200 ppm has been shown more ability to prevent the
formation of free radicals than industrial antioxidants [69].
And also according to other studies Jang-HyukAhn and Young-Pil Kim found after
adding herbal extracts (rosemary, broccoli sprout and citrus) to sunflower oil, these
extracts have the potential to prevent the oxidation of fat found in sunflower oil [70].
1.4.2 Using the sage extract as a natural antioxidant for vegetable oil
Some previous studies have indicated that sage and rosemary contain similar types
of phenolic compounds. Antioxidative activity attributed to their contain on rosmarinic
acid and carnosic .
Other studies have indicated that sage contain more active compounds such as
phenolic acids, flavonoids and terpenoids [71].
Relative to rosemary, sage is especially prolific in the outputting of phenolics and
other flavonoids derivatives, particularly those structurally regarding the rosmarinic acid.
Some previous studies have indicated that rosmarinic acid is responsible for
antioxidant activity of sage [72].
In general, sage produces lower amount of phenolics ,carnosic acid and other
compounds than does rosemary [40].
Abdalla and Roozen found through experiments that sunflower oil contains 570
ppm natural α-tocopherol added to it sage extract showed high antioxidant activity through
secondary and primary oxidation [73].
Salvia officinalis is a kind of sage its extract has a high ability to radical scavenging.
Sage extract had almost 2 times the amount of diterpenoidphenolics less than
rosemary extract, and about 2.7 the level of carnosic acid and carnosol.
Many of the researches indicated that carnosic acid is powerful antioxidant in
rosemary.
26
This is the main reason that sage extract is less used than rosemary as an antioxidant
[43].
Figure 7 -Some anti-oxidant compounds are present in sage
27
1.4.3 Using the thyme extract as a natural antioxidant for vegetable oil
This type belongs to the aromatic herbs that are usually added to foods to give flavor
and aroma.As a spice it can be used dry or fresh leaves [74].
Thyme contains rosmarinic acid eriodyctiol, luteolinglycosides,luteolin, and
glycuronids of apigenin which are the major phenolic compounds.
Several types of thyme such as mastichina, caespititius and camphorate are showed
antioxidant activity similar to BHT and α-tocopherol [40].
Thyme extract whichis added at concentration of 1%to sunflower oil prevent
oxidation operation through its storage for 29 days in different temperature conditions
[75].
Turek and Stintzing indicated that thyme extracts has an antioxidant activities
which were determined by measuring the pv (peroxide value) which are present in in
sunflower oil at 60 ° C and found that when adding thyme extract at concentration 6000
mg/kg oil to sunflower oil shown effective as an antioxidant almost equal1000 mg
BHA/kg oil that were added to the same oil [76].
Many of the researches indicated that essential oil of this herb contents high of
carvacrol and thymol which have strong antioxidant activity [77].
1,8-cineole, α-terpineol,carvacrol, and thymol considered as the mainly aroma
compounds found in thyme.
Thymol is the highest effective antioxidative compounds and is considered as the
mainly aroma component in thyme [78].
Thymol and carvacrol each one of them has1 aromatic ring with1 −OH group,pcymene contains1 aromatic group and 1-terpineol contains1 −OH group, existence of
aromatic ring and OH groups which is responsible for the anti-oxidant properties of these
compounds [40].
28
Figure 8 -Some anti-oxidant compounds are present in thyme
1.4.4 Using the ginger extract as a natural antioxidant for vegetable oil
This type of spices was added to foods to give flavor [79] and it also possesses the
properties of anti-oxidant materials [80]. These properties are because it contains
gingeronesshogaols and gingerols.
29
Dried and fresh ginger contain high amounts of the camphene, volatile oils , pcineole, zingiberene, pentadecanoic acid and alpha-terpineol [81].
Zingiberofficinale as a type of ginger contains several antioxidants such as, ascorbic
acid, terpenoids, as terpenoids beta-carotene, alkaloids, flavones, flavonoids, rutin, and
glycosides. Because of its high content of antioxidants, it can be considered main source
of natural antioxidants [82, 83].
Brewer the researcher concluded that ginger extract when added to sunflower oil
the resultsshown that its antioxidant activity was nearly too synthetic antioxidants (BHT
and BHA).Antioxidative activity of compounds which separated from ginger such as
8diarylheptanoids and 5 compounds related to ginger shown higher than α-tocopherol[40].
Figure 9 -Some anti-oxidant compounds are present in ginger
30
Salariya and Habib the researchers concluded that the solution obtained from ginger
which was added to sunflower oil which storage for 60 days appeared good thermal
stability [84].
When ginger extract was added with concentrations 1600 and 2400ppm to
sunflower oil, the results showed that the process of out-oxidation was largely prevented
and there is no difference between using ginger extract with 1600 and 2400ppm and
synthetic antioxidants with 200 ppm [85].
Some previous studies have indicated that effectiveness of antioxidants BHA at
2400 and1600 ppm were less effective than the effective of ginger extract but it is almost
equal to efficiency of BHT at 2400ppm when added to sunflower oil [84].
1.4.5 Using the oregano extract as a natural antioxidant for vegetable oil
This type of herb belongs to the Lamiaceaefamily [86].
Because it contains compounds such carvacrol, thymol, c-terpinene, linalol, pcymene, and sesquiterpenes and other monoterpenes. This makes it possess anti-oxidant
and anti-bacterial properties [87].
Origanum vulgare ssp and hirtum,origanum and P. longiflora are types of oregano
which contain high phenolic compounds. Antioxidant efficacy of tocopherol is less
effective of these compounds.
Hydroxycinnamic acid and rosmarinicacid ascompounds are present at high
concentrations in oregano extracts and have a high antioxidant activity [88, 89].
Some previous studies have indicated that the essential oil which was extracted
from oregano contain thymol and carvacrol. Perhaps these substances are responsible for
the antioxidant effect of oregano [90, 91].
31
Figure 10 -Some anti-oxidant compounds are present in Oregano
Oregano extract which was extracted by using acetone is highly active in sunflower
oil in comparison with 20 % oil-in-water emulsion through oxidation at 60 °C in the dark
[92].
Jorge and Veronezi found that oregano showed a high effect in preventing the
formation of polar compounds compared with antioxidant TBHQ. In addition, when added
it at 3,000 mg/kg concentration, antioxidant protection has been shown to be better than
industrial antioxidants [93].
32
CHAPTER
2
OBJECTS AND METHOD OF RESEARCH
2.1 Research objectives
-Conducting a process of extraction of herbs used (thyme, ginger, rosemary, sage)
by the use of ethanol at a concentration of 95% and 70%.
-Conducting a process of extraction of herbs used (thyme, ginger, rosemary, sage)
by the use of (ethanol 95%, propylene glycol 95%) and propylene glycol at a concentration
of 95%.
-Determination of the content of polyphenols for herbal extracts (thyme, ginger,
rosemary, sage).
-Measuring the free radical scavenging activity (DPPH) of herbal extracts (thyme,
ginger, rosemary, sage).
-Measure both the peroxide value and the acid value in sunflower oil after adding
each extract obtained from the herb used every 15 days for two months.
-Measure both the peroxide value and the acid value in sunflower oil after adding a
mixture of extract obtained from the herbs used every 15 days for two months.
-Comparison between the peroxide values and acid values after the addition of each
single herb extract and the addition of a mixture of herb extract.
- Comparison between the effectiveness of herbal extracts and the effectiveness of
industrial antioxidants on the process of oxidation of sunflower oil when adding each
extract separately.
- Comparison between the effectiveness of herbal extracts and the effectiveness of
the industrial antioxidant on the process of oxidation of sunflower oil when adding a
mixture of extracts from these herbs.
-Comparison between the effect of adding herb extracts separately and adding a
mixture of extracts to sunflower oil on the oxidation process of sunflower oil.
33
2.2 The preparation of herbs extracts using ethanol 95%
Materials and equipment used:
- Dried herbs (thyme, ginger, rosemary, sage).
- Solvent ethanol 95%.
-Filter paper in order to filter the herb extract.
-Balance for weighing samples.
Conduct extraction: dried herbs (thyme, ginger, rosemary , sage) used .The grinding
of the leaves was done using a small electric mill and was put the leaves powder in small
glass containers until extraction. Then 5 grams of each sample was weighed. Then they
were put in erlenmeyer flasks .Then 100 ml of solvent ethanol 95%were added to the herbs
samples, and direct extraction was performed at the boiling point of the ethanol for 6
hours.
Then the centrifuged process was performed for 10 minutes.
The filtration process was done for sample using filter paper. Then the extract
obtained from the herb was placed in the fridge until usage.
When conducting the first experiment on herbal extracts separately. The solution
containing each sample of herbal extract was added to 100 ml of sunflower oil at a
concentration of 1%.
When conducting the second experiment on a mixture of herbal extracts, a mixture
of extracts (thyme- ginger, rosemary- thyme, ginger- rosemary, sage- rosemary) was
added at concentration 1% to 100 ml of refined sunflower oil sample.
2.3 The preparation of herbs extracts using ethanol 70%
Materials and equipment used:
- Dried herbs (thyme, ginger, rosemary, sage).
- Solvent ethanol 70%.
34
-Filter paper in order to filter the herb extract.
-Balance for weighing samples.
Conduct extraction: dried herbs (thyme, ginger, rosemary , sage) used .The grinding
of the leaves was done using a small electric mill and was put the leaves powder in small
glass containers until extraction. Then 2 grams of each sample was weighed. Then they
were put in erlenmeyer flasks .Then 15 ml of solvent ethanol 70%were added to the herbs
samples. The mixture was then placed in a water bath at 70 ° C for 30 min. The filtration
process was done for sample using filter paper. Then the extract obtained from the herb
was placed in the fridge until usage.
When conducting the first experiment on herbal extracts separately. The solution
containing each sample of herbal extract was added to 100 ml of sunflower oil at a
concentration of 1%.
When conducting the second experiment on a mixture of herbal extracts, a mixture
of extracts (thyme- ginger, rosemary- thyme, ginger- rosemary, sage- rosemary) wasadded
at concentration 1% to 100 ml of refined sunflower oil sample.
2.4 The preparation of herbs extracts using propylene glycol 95%
Materials and equipment used:
- Dried herbs (thyme, ginger, rosemary, sage).
- Solvent propylene glycol 95%.
-Filter paper in order to filter the herb extract.
-Balance for weighing samples.
-Oven for solvent evaporation.
Conduct extraction: dried herbs (thyme, ginger, rosemary , sage) used .The grinding
of the herbs was done using a small electric mill and was put the leaves powder in small
glass containers until extraction. Then 10 grams of each sample was weighed. Then they
35
were put in erlenmeyer flasks, and then 100 ml of solvent propylene glycol 95% was
added to the herbs samples .The mixture was left at room temperature for one day .The
filtration process was done for the mixture using filter paper. The solvent was evaporated
by placing it in the oven at 40 ° C. Then the extract obtained from the herb was placed in
the fridge until usage.
When conducting the first experiment on herbal extracts separately. The solution
containing each sample of herbal extract was added to 100 ml of sunflower oil at a
concentration of 1%.
When conducting the second experiment on a mixture of herbal extracts, a mixture
of extracts (thyme- ginger, rosemary- thyme, ginger- rosemary, sage- rosemary) wasadded
at concentration 1% to 100 ml of refined sunflower oil sample.
2.5 The preparation of herbs extracts using propylene glycol 95% and ethanol
95%
Materials and equipment used:
- Dried herbs (thyme, ginger, rosemary, sage).
- A mixture of solvents (50 ml ethanol 95% and 50 ml propylene glycol 95%(.
-Filter paper in order to filter the herb extract.
-Balance for weighing samples.
-Oven for solvent evaporation.
Conduct extraction: dried herbs (thyme, ginger, rosemary , sage) used .The grinding
of the leaves was done using a small electric mill and was put the leaves powder in small
glass containers until extraction. Then 10 grams of each sample was weighed into one
erlenmeyer flasks, and then 100 ml of a mixture of solvents (50 ml ethanol 95% and 50
ml propylene glycol 95%) was added to the herbs samples .The mixture was left at room
temperature for one day .The filtration process was done for the mixture using filter paper.
36
The solvent was evaporated by placing it in the oven at 40 ° C. Then the extract obtained
from the herb was placed in the fridge until usage.
When conducting the first experiment on herbal extracts separately. The solution
containing each sample of herbal extract was added to 100 ml of sunflower oil at a
concentration of 1%.when conducting the second experiment on a mixture of herbal
extracts, a mixture of extracts (thyme- ginger, rosemary- thyme, ginger- rosemary, sagerosemary) was added at concentration 1% to 100 ml of refined sunflower oil sample.
2.6 Determination of the content of polyphenols for herbs extracts
Materials and equipment used:
-Herbal extracts (thyme, ginger, rosemary , sage) .
- Folin–Ciocalteu phenol reagents.
- Gallic acid: prepare a standard series of Gallic acid with concentrations (10, 20,40,
60, 80 mg /ml).
-Sodium carbonate: A solution of sodium carbonate is prepared at a concentration
of 7.5%.
-Spectrophotometer device to measure the absorption of samples.
Conduct the experiment: polyphenol content was determined for herbal extracts
used according to the Folin–Ciocalteu method .By adding 0.5 ml of each extract from the
herbs to 10 ml of distilled water and then adding 2.5 ml of (Folin–Ciocalteu phenol
reagents). After five minutes 2 ml of 7.5 %sodium carbonate solution was added to the
previous mixture .The solution is then left in the dark for sixty minutes.
The absorbance of the solution is then measured using the spectrophotometer at
750nm and the results were expressed as mg of Gallic acid equivalent per ml [97].
37
After we dissolve 0.1 mg of Gallic acid in distilled water in a 100 mL calibrated
balloon and prepare a standard series of Gallic acid at concentrations (10,20,40, 60, 80
mg / ml) and complete the volume with distilled water up to 100mL.
Then, the absorbance is measured at a 750 nm for the samples of Gallic acid and
drew the calibration curve of the Gallic acid.
2.7 Measuring the free radical scavenging activity (DPPH) of herbal extracts
Materials and equipment used:
- Herbs extracts (thyme, ginger, rosemary , sage) .
- DPPH) 1,1-diphenyl-2-picrylhydrazyl(.
-Spectrophotometer device to measure the absorption of samples.
-Methanol.
Conduct the experiment: the determining of total free radical scavenging capacity
of extracts from different plant samples was estimated according to the following method.
A solution of radical is prepared by adding 2.4 mg DPPH in 100 mL methanol. Test
solution) 0.5μl of herbs extract (was added to 3.995 ml of methanol- DPPH. The mixture
was vigorously shaken and kept in the dark at room temperature for 30 minutes.
Absorption was measured using a spectrophotometer at 515nm.
Absorption was also measured for the same solution) DPPH (but without the
presence of the sample blank.
The measurement was done three times per sample [98].
The capability to scavenge the DPPH radical was calculated using the following
equation:
DPPH Scavenged (%) = ((AB–AA)/AB) ×100
Where is AB - is absorbance of solution but without the presence of the sample.
38
AA - absorbance of solution of the sample.
2.8 Determine the peroxide value of oil samples
The peroxide value was determined according to GOST Р 51487-99[99].
Preparation of solutions:
-A solution of potassium iodide is prepared at concentration 55%.
-The starch solution is prepared as follows: 5 g of a soluble starch is mixed with 30
cm of water, add this mixture to 1000 mm boiling water and boil for 3 minutes.
-A solution of sodium thiosulfate with a molar concentration of 0.1 M.
Conduct the experiment: in a flask with a 5 g of sample, add 10 cm of chloroform,
rapidly dissolve the sample, add 15 cm of acetic acid and 1 cm of 55% solution of
potassium iodide, after that the flask is immediately closed, the contents are stirred for 1
minute and left for 5 minutes in a dark place at a temperature of 15-25 ° C.
Add 75 ml of distilled water to the flask, carefully stir and add the starch solution
until we see weak homogeneous violet-blue color.
Isolated iodine is titrated with a solution sodium thiosulfate to milky-white color,
stable for 5 seconds.
The peroxide value was calculated using the following formula:
𝑥=
1000(𝑣 − 𝑣0)𝑐
𝑚
Where is V- the volume of sodium thiosulfate solution used for oil sample
definition, mm.
V0 -The volume of sodium thiosulfate solution used for control definition, mm.
c -The actual concentration of the used thiosulfate sodium solution.
m -Weight of the sample of the product, g.
39
2.9 Determine the acid value of oil samples
The acid value was determined according to GOST R 50457-92 [100].
Materials used:
-Refined sunflower oil.
-Phenolic phenols: weighs 1 gram of phenolic phenols and melts in 100 ml of
ethanol.
- Potassium hydroxide: carefully weigh5.61 grams of sodium hydroxide and place
in a 1000 ml flask. Then the distilled was added until the mark.
-Ether.
Conduct the experiment: in a conical flask with a capacity of 250 mm, 5 g of
sunflower oil was weighed. Then add 50 mm of ether to the sample. The contents of the
flask are stirred by shaking the flask by hand. If the oil does not dissolve, it is heatedto
(50) ° C in a water bath, then cooled to 20 ° C. A few drops of phenolphthalein are added
to the solution. Then the solution is calibrated with a solution of potassium hydroxide at
a concentration 0.1 N until a slightly pink color is obtained and stable for 30 s.
The acid value was calculated using the following formula:
AV=
𝑣.56,1.𝑐
m
Where is 𝑣 - volume of a solution of potassium hydroxide in a molar concentration
(KOH) = 0.1 mol / dm3, expended on titration, mm.
C - The actual concentration of potassium hydroxide.
56.1 - Is the KOH mass in 1 cm of the molar concentration solution (KOH) = 0.1mol
/ dm3 (0.1 N).
m - Weight of the sample, g.
40
CHAPTER
3 RESULTSAND DISCUSSION
3.1 The amount of phenolic compounds for herbs extracts
The amount of phenolic compounds was determined by the linear regression
equation which is obtained from the calibration curve of Gallic acid as a standard y =
0.009x + 0.0768 ;R² = 0.9869 based on the Gallic acid percentage. In the regression
equation of the calibration curve of Gallic acid, Y is the absorption rate that was read at a
wavelength of 765 nm and X is the concentration of phenolic compounds based on mg
gallic acid/mL.
0.9
Absorption (765 nm)
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
20
40
60
80
Concentration (mg/ml)
Figure 11 - Standard curve of Gallic acid
41
100
The amount of phenolic compounds for various extracts ranged from 11.12 to
45.17. The Total polyphenol content of the extracts can be ranked in the order rosemary
(45.17)>thyme (36.15) > sage (25.74) > ginger (11.12).The results show that the rosemary
extract contains the highest value of polyphenols.
Table 4 – The amount of phenolic compounds for various extracts extracted with
ethanol 95%
Herbs
Rosemary
Thyme
Ginger
Sage
Total polyphenol content (mg Gallic acid/mL )
45.17 mg/ml
36.15 mg/ml
11.12 mg/ml
25.74 mg/ml
3.2 The DPPH radical scavenging ability of the herbs extracts
The DPPH is a stable radical with a maximum absorption at 517 nm that can readily
undergo scavenging by antioxidant. It has been widely used to evaluate the antioxidative
activity of plant extracts .All the extracts showed different levels of DPPH radical
scavenging activity.
Table 5 – DPPH free radical scavenging activity of different extracts from herbs
extracted with ethanol 95%
Herbs
inhibition DPPH radical%
Rosemary
85.15
Thyme
76.55
Ginger
36.44
Sage
55.64
The results show that the DPPH radical scavenging ability of the extracts can be
ranked in the order rosemary (85.15%) >thyme (76.55%) > sage (55.64 %) > ginger
(36.44%). The observed differential scavenging activities of the extracts against the DPPH
system could be due to the presence of different compounds in the extract.
42
3.3 The peroxide values and acid values of refined sunflower oil samples
3.3.1 Effect of adding herb extracts extracted with ethanol 95% separately or
BHT on the peroxide values and on acid values of refined sunflower oil samples
In this experiment, extracts were added separately at concentration 1% to 100 ml of
refined sunflower oil sample and BHT at concentration 180 mg / Kg.
Initially, the peroxide value of refined sunflower oil was determined without any
addition and the result was 3.66 meq/kg.
Every 15 days for two months the peroxide value of the oil samples containing the
plant extracts or the industrial antioxidant was measured and the results shown in the table
6.
Table 6 – The effects of (BHT), thyme extract, ginger extract, rosemary extract,
added to the refined sunflower oils, on peroxide values
Days
The
The sample
The sample of
The sample of
The sample
The
sample of
of sunflower
sunflower oil
sunflower oil
of sunflower
sample
sunflower
oil with
with rosemary
with
oil with sage
of
oil
thyme
extract
ginger
extract
sunflower
without
extract
extract
oil with
additions
BHT
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.74
3.72
3.75
3.77
3.79
30days
5.3
3.84
3.82
3.88
3.89
3.98
45days
5.95
3.92
3.91
4.03
4.05
4.3
60days
6.5
4
3.97
4.012
4.015
4.4
Through the results, we note that the values of peroxide of the sample of sunflower
oil without any additions have increased from 3.66 to 6.5.While the oil samples- whether
43
containing the herbal extracts or the industrial antioxidant- the increase in the values of
peroxide were few. The results were represented on a diagram showing the changes in the
peroxide values of the various oil samples.
7
Peroxide value )meq/kg(
6
5
4
3
2
1
0
1
15
30
45
60
Storage time (day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 12 - Effect of adding BHT and herbal extracts extracted with ethanol 95%
on peroxide value of sunflower oil
In comparing the different samples, we note that the lowest peroxide value was for
the sample of sunflower oil containing the rosemary extract. This is due to the high
percentage of polyphenols. On the other hand, the peroxide values of all samples
44
containing herb extracts were lower than the peroxide value of the oil sample containing
the industrial antioxidant.
Thus, we conclude that herbal extracts have a higher effectiveness in preventing the
oxidation of sunflower oil than the industrial antioxidant.
First, the acid value of the refined sunflower oil was determined without any
additives and the result was 0.114 mg КОН / g.
The acid value of refined sunflower oil samples- whether free of additives or
containing (BHT) or the herbal extracts added separately at concentration 1% to the oilwere determined every 15 days for two months, and the results were shown in table 7.
Table 7 – The effects of (BHA), thyme extract, ginger extract, rosemary extract,
added to the refined sunflower oils, on its acid values
Days
The
The sample
The
The sample of
The sample
The
sample of
of sunflower
sample of
sunflower oil
of sunflower
sample of
sunflower
oil with
sunflower
with
oil with sage
sunflower
oil
thyme
oil with
ginger
extract
oil with
without
extract
rosemary
extract
additions
BHT
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.2
0.18
0.22
0.24
0.24
30days
0.47
0.28
0.24
0.32
0.34
0.36
45days
0.65
0.33
0.3
0.41
0.43
0.47
60days
0.87
0.4
0.38
0.48
0.53
0.58
From the results, we can observe the increase in the acidity value of the sample of
refined sunflower oil, which is free of any additives, was big.
But other oil samples whether containing (BHT ) or the herbal extracts added
separately to the oil the change in acidity values was small.
45
The results were represented on a diagram showing the changes in the acid values
of the various oil samples.
1
0.9
Acid value ) mg КОН / g(
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 13 – Effect adding of BHT and herbal extracts extracted with ethanol
95%on acid value of sunflower oil
After comparing the different samples, we note that the sample with the lowest acid
value is the sample containing the rosemary extract, because it contains a higher
proportion of polyphenols than other samples.
46
Furthermore, the acid value of the oil sample containing the industrial antioxidant
was greater than the acid values of all samples containing herb extracts.Thus, we conclude
that the industrial antioxidant has a lower effectiveness in preventing the oxidation of
sunflower oil than herbal extracts.
3.3.2 Effect of adding a mixture of herb extracts extracted with ethanol 95%
or BHT on the peroxide values and on acid values of refined sunflower oil samples
In this experiment, a mixture of extracts (thyme- ginger, rosemary- thyme, gingerrosemary, sage- rosemary) was added at concentration 1% to 100 ml of refined sunflower
oil sample and BHT at concentration 180 mg / Kg.
In the beginning, the peroxide value was measured for the refined sunflower oil
sample, which did not contain any additives, and the result was 3.66 meq/kg. The peroxide
values were then measured for different oil samples containing either a mixture of herb
extracts or synthetic antioxidant every 15 days for two months. The results were as follows
in table 8.
Table 8 – Effect of adding a mixture of herbal extracts or synthetic antioxidants on
the peroxide values of refined sunflower oil.
Days
The
The
The sample of
The sample of
The sample
The
sample of
sample of
sunflower oil
sunflower oil
of sunflower
sample of
with
oil with sage
sunflower
sunflower sunflower with rosemary
oil
oil with
and thyme
ginger and
and rosemary
oil with
without
thyme
extract
rosemary
extract
BHT
additions
and
extract
ginger
extract
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.72
3.70
3.74
3.75
3.79
47
30days
5.3
3.81
3.76
3.83
3.86
3.98
45days
5.95
3.9
3.83
3.92
4.01
4.3
60days
6.5
3.96
3.88
4.02
4.10
4.4
After examining the results we noted, an increase in the peroxide value of the
sample of sunflower oil that does not contain additives was larger than other samples.
On the other hand, the peroxide values of refined sunflower oil samples containing
either a mixture of herb extracts or synthetic antioxidant have not changed
significantly.The results in the table were represented on figure 14 showing the increase
in peroxide values for all oil samples.
Peroxide value )meq/kg(
7
6
5
4
3
2
1
0
1
15
30
45
60
Storage time (day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 14 -Effect of adding BHT and a mixture of herbal extracts extracted with
ethanol 95% on peroxide value of sunflower oil
The comparison between the different samples shows that the sample containing
the rosemary-thyme extract had the lowest peroxide value. The peroxide value for this
48
sample (3.88 meq/kg) was less than the limit of acceptable values (10 meq/kg), stipulated
by the standard GOST 1129-2013 for sunflower oil [101] because, rosemary and thyme
extract contained the highest concentration of phenolic substances. It was also found that
all the samples of oil containing a mixture of herbal extracts have lower peroxide values
than the sample containing the synthetic antioxidant.
We can conclude that the mixture of herbal extracts has a greater effectiveness in
delaying the occurrence of the process of oxidation of refined sunflower oil than the
synthetic antioxidant.
At first, the acid value was measured for the refined sunflower oil sample, which
did not contain any additives, and the result was as follows 0.114 mg КОН / g.
Every 15 days for two months, the acid value of the oil samples containing a mixture
of herbal extracts or the industrial antioxidant was measured and the results were as
follows in the table 9.
Table 9 – Effect of adding a mixture of herbal extracts or industrial antioxidants on
the acid values of refined sunflower oil.
Days
The
The
sample of sample of
The sample of
The sample of
The sample
The
sunflower oil
sunflower oil
of
sample of
with
sunflower
sunflower
sunflower sunflower with rosemary
oil
oil with
and thyme
ginger and
oil with
oil with
without
thyme
extract
rosemary
sage and
BHT
additions
and
extract
rosemary
ginger
extract
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
0.31
0.19
0.17
0.21
0.22
0.24
30days
0.47
0.25
0.22
0.3
0.34
0.36
45days
0.65
0.31
0.27
0.38
0.4
0.47
15
days
49
60days
0.87
0.37
0.31
0.44
0.51
0.58
Through the results, we note that the acid value of the sample of sunflower oil
without any additions have increased from 0.114 to 0.87.However, the oil samples,
whether containing the synthetic antioxidant or the mixture of herbal extracts, the increase
in the values of acid were few. The results in the table were represented in figure 15
showing the increase in acid values for all oil samples.
1
Acid value ( mg КОН / g)
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
10
20
30
40
50
60
70
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme
extract
sunflower oil sample with ginger and rosemary
extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 15 -Effect of adding BHT and a mixture of herbal extracts extracted with
ethanol 95% on acid value of sunflower oil
50
Table 10 – Regression equations describing the change in acid value for the samples
of sunflower oil within two months
The sample of sunflower oil
The regression equation
Coefficient of
approximation
The sample of sunflower oil without
additions
y = 0.0125x + 0.1049
R² = 0.9972
The sample of sunflower oil with thyme and
ginger extract
y = 0.0043x + 0.1179
R² = 0.9959
The sample of sunflower oil with rosemary
and thyme extract
y = 0.0033x + 0.1165
R² = 0.9954
The sample of sunflower oil with ginger and
rosemary extract
y = 0.0055x + 0.1212
R² = 0.9908
The sample of sunflower oil with sage and
rosemary extract
y = 0.0066x + 0.1186
R² = 0.9902
The sample of sunflower oil with BHT
y = 0.0078x + 0.1158
R² = 0.9981
y-Acid value ( mg КОН / g);x- Storage time(day).
After comparing the different samples, we note that the sample with the lowest acid
value is the sample containing the rosemary-thyme extract.
The acid number for this sample (0.31 mg KOH / g of oil) was less than the limit
of acceptable values (0.4 mg KOH / g of oil), stipulated by the standard GOST 1129-2013
for sunflower oil, because they contain compounds such as rosmanol, rosmaridiphenol,
and carnosol which are effective as antioxidants.
After adding both herbal extracts and industrial antioxidants to refined sunflower
oil samples, the sample containing the industrial antioxidant has the highest acid value
among other samples containing herbal extracts.
Therefore, we conclude that the synthetic antioxidant has a less effectiveness in
delaying the occurrence of the process of oxidation of refined sunflower oil than a mixture
of herbal extracts.
51
3.3.3 Effect of adding herb extracts extracted with ethanol 70% separately or
BHT on the peroxide values and on acid values of refined sunflower oil samples
In this experiment, extracts were added separately at concentration 1% to 100 ml of
refined sunflower oil sample and BHT at concentration 180 mg / Kg.
At the first day, the peroxide value of the oil sample was determined without any
additions and the value was 3.66 meq/kg. After every 15 days for 60 days, the peroxide
values of the oil samples, whether containing the synthetic antioxidant or herb extracts,
was measured and the results were as in the table 11.
Table 11 – Effect of adding herbal extracts or BHT on the peroxide values of refined
sunflower oil
Days
The
The sample
The sample of
The sample of
The sample
The
sample of
of sunflower
sunflower oil
sunflower oil
of sunflower
sample of
sunflower
oil with
with rosemary
with
oil with sage
sunflower
oil
thyme
extract
ginger
extract
oil with
without
extract
extract
BHT
additions
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.74
3.72
3.76
3.78
3.79
30days
5.3
3.86
3.83
3.89
3.9
3.98
45days
5.95
3.97
3.94
4.05
4.07
4.3
60days
6.5
4.09
3.99
4.14
4.15
4.4
Among all oil samples, refined sunflower oil sample without any additives showed
the highest increased in peroxide value. When comparing different samples, the increased
of peroxide value during 60 days storage were small for oil samples containing the herbal
extracts or BHT. The results were represented on the following chart.
52
7
Peroxide value )meq/kg(
6
5
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 16 -Effect of adding BHT and herbal extracts extracted with ethanol 70%
on peroxide value of sunflower oil
We note from the results that the oil sample containing the rosemary extract had the
lowest peroxide value, due to their high content of polyphenols.
53
The results show that after the addition of plant extracts to refined sunflower oil
samples, the peroxide values of these samples were lower than the value of peroxide when
adding BHT.
Thus, we conclude that the BHT antioxidant had the potential to inhibit the
oxidation process of refined sunflower oil less than herbal extracts.
At the beginning of the experiment, the acid value of the refined sunflower oil
sample was measured; it was0.114 mg КОН / g. After the addition of BHT antioxidant or
the herb extracts to the refined sunflower oil samples, the acid values of these samples
were determined every 15 days for 60 days.
Table 12 – The effect of herbal extracts or industrial antioxidants on the acid values
of refined sunflower oil when each was added separately
Days
The
The
The
The sample
The sample
The
sample of
sample of sample of
of
of sunflower
Sample of
sunflower
sunflower sunflower
sunflower
oil with sage
sunflower
extract
oil with
oil
oil with
oil with
oil with
without
thyme
rosemary
ginger
additions
extract
extract
extract
BHT
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.2
0.19
0.22
0.24
0.24
30days
0.47
0.31
0.27
0.31
0.33
0.36
45days
0.65
0.39
0.33
0.43
0.44
0.47
60days
0.87
0.47
0.39
0.52
0.54
0.58
From the results, we note that the sample that has the highest acid value after 60
days is the sample of oil free of additives.
From the results in table 12 we note that the increased of acid value during 60 days
storage were small for oil samples containing the herbal extracts or BHT. These results
were represented on the chart.
54
1
Acid value ) mg КОН / g(
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 17 -Effect of adding BHT and herbal extracts extracted with ethanol
70%on acid value of sunflower oil
Among all oil samples, refined sunflower oil sample with rosemary extract showed
the lowest increase in acid value. This may be because it contains more polyphenols than
other samples. The increase in the acid values of the oil samples containing the herb
extract is less than the increase in the acid value of the oil sample containing the synthetic
antioxidant.
Therefore, we conclude that herbal extracts have the potential to inhibit the
oxidation process of refined sunflower oil more than the industrial antioxidant.
55
3.3.4 Effect of adding a mixture of herb extracts extracted with ethanol 70%
or BHT on the peroxide values and on acid values of refined sunflower oil samples
In this experiment, a mixture of extracts (thyme- ginger, rosemary- thyme, gingerrosemary, sage- rosemary) was added at concentration 1% to 100 ml of refined sunflower
oil sample and BHT at concentration 180 mg / Kg.
On the first day, the peroxide value of oil sample without additions was measured
and the value was 3.66 meq/kg. Every 15 days for two months, the peroxide value of the
oil samples containing the plant extracts or the synthetic antioxidant was measured and
the results were as in the table 13.
Table 13 – Effect of adding a mixture of herbal extracts or industrial antioxidants
on the peroxide values of refined sunflower oil
Days
The
The
The sample
The sample of
The
The
sample of
sample of
of sunflower
sunflower oil
sample of
Sample of
oil with
with
sunflower
sunflower
sunflower sunflower
oil
oil with
rosemary and
ginger and
oil with
oil with
without
thyme
thyme
rosemary
sage and
BHT
additions
and
extract
extract
rosemary
ginger
extract
extract
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.72
3.71
3.74
3.75
3.79
30days
5.3
3.83
3.78
3.86
3.88
3.98
45days
5.95
3.92
3.86
3.99
4.02
4.3
60days
6.5
3.98
3.92
4.09
4.12
4.4
A sample of refined sunflower oil free from herb extracts or BHT increased its
peroxide value more than other samples as it increased from3.66 to6.5.
56
The peroxide values of the oil samples did not change much after 60 days except
for the oil sample free from any additives; the results were represented on the following
chart.
7
Peroxide value )meq/kg(
6
5
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 18 -Effect of adding BHT and a mixture of herbal extracts extracted with
ethanol 70% on peroxide value of sunflower oil
From the results, we note that, the sample containing the rosemary-thyme extract
has the lowest peroxide value among the other samples. The peroxide value for this sample
(3.92 meq/kg) was less than the limit of acceptable values (10 meq/kg), stipulated by the
standard ТР ТС 024/2011 for technical regulations for oil and fat products [102].
57
This may be because they contain a higher percentage of compounds that are
effective as antioxidants.
By observing the results in table 13, we observe that the increase in the peroxide
values of all the oil samples containing the herb extracts is less than the increase in the
peroxide value of the oil sample containing the industrial antioxidant.
Thus we can conclude that herbal extracts that have been added to sunflower oil
have the potential to delay the occurrence of oxidation of oil more than the industrial
antioxidant.
At first, the acid value was measured for the refined sunflower oil sample, which
did not contain any addition, and the result was as follows 0.114 mg КОН / g.
After adding a mixture of herbal extracts and BHT to the oil samples and measuring
the acid values for each sample after 15 days and for two months. The results were as in
the table14.
Table 14 – Effect of adding a mixture of herbal extracts or industrial antioxidants
on the acid values of refined sunflower oil
Days
The
The
The
The sample of
The
The
sample of
sample of sample of
sunflower oil
sample
Sample of
sunflower
sunflower sunflower
with
of
sunflower
oil
oil with
oil with
ginger and
sunflower
oil with
without
thyme
rosemary
rosemary
oil with
BHT
additions
and
and
extract
sage and
ginger
thyme
rosemary
extract
extract
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.2
0.19
0.23
0.24
0.24
30days
0.47
0.26
0.24
0.32
0.34
0.36
45days
0.65
0.32
0.29
0.42
0.44
0.47
60days
0.87
0.38
0.34
0.49
0.51
0.58
58
In the table we note that the change in the value of the acid of the sample of the oil
containing no addition is much greater compared to the change in the values of acid of the
oil samples, whether containing a mixture extracts of herbs or industrial antioxidant.
However, the oil samples, whether containing the synthetic antioxidant or the
mixture of herbal extracts, the increase in the values of acid were few. The change in the
peroxide values was represented on the chart.
1
0.9
Acid value ) mg КОН / g(
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
10
20
30
40
50
60
70
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 19 - Effect of adding BHT and a mixture of herbal extractsextracted with
ethanol 70% on acid value of sunflower oil
59
Table 15 – Regression equations describing the change in acid value for the samples
of sunflower oil within two months
The sample of sunflower oil
The regression equation
Coefficient of
approximation
The sample of sunflower oil without
additions
y = 0.0125x + 0.1049
R² = 0.9972
The sample of sunflower oil with thyme and
ginger extract
y = 0.0044x + 0.1219
R² = 0.9914
The sample of sunflower oil with rosemary
and thyme extract
y = 0.0037x + 0.1223
R² = 0.9885
The sample of sunflower oil with ginger and
rosemary extract
y = 0.0064x + 0.1228
R² = 0.9917
The sample of sunflower oil with sage and
rosemary extract
y = 0.0067x + 0.1266
R² = 0.9881
The sample of sunflower oil with BHT
y = 0.0078x + 0.1158
R² = 0.9981
y-Acid value (mg КОН / g); x- Storage time (day).
The results of the experiment showed that the sample that has the least acidity value
is the oil sample container of the rosemary-thyme extract.
This may be because they contain a higher percentage of polyphenols than other
samples.
The acid number for this sample (0.34 mg KOH / g of oil) was less than the limit
of acceptable values (0.6 mg KOH / g of oil), stipulated by the standard ТР ТС 024/2011
for technical regulations for oil and fat products.
From the results in table 14 we note that increase in the acid value of the oil sample
containing BHT is greater than the increase in the acid values of all the oil samples
containing the herb extracts.
Therefore, we can conclude that the industrial antioxidant that added to sunflower
oil has the potential to delay the occurrence of oxidation of oil less than herbal extracts.
60
3.3.5 Effect of adding herb extracts extracted with ethanol 95%and propylene
glycol 95% or BHT on the peroxide and on acid values of refined sunflower oil
samples
In this experiment, extracts were added separately at concentration 1% to 100 ml of
refined sunflower oil sample and BHT at concentration 180 mg / Kg.
Firstly, the peroxide value of refined sunflower oil was determined without any
addition and the value was 3.66 meq/kg.
Then the herbal extracts and the industrial antioxidant were added separately to the
oil samples. Peroxide values were measured for each sample after 15 days and for two
months. The results were as follows in the table16.
Table 16 – The effects of (BHA), thyme extract, ginger extract, rosemary extract,
added to the refined sunflower oils, on peroxide values respectively were
Days
The
The sample
The sample of
The sample
The sample
The
sample of
of sunflower
sunflower oil
of sunflower
of
sample of
sunflower
oil with
with rosemary
oil with
sunflower
sunflower
oil
thyme
extract
ginger
oil with
oil with
without
extract
extract
sage
BHT
additions
extract
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.76
3.74
3.77
3.78
3.79
30days
5.3
3.88
3.85
3.91
3.93
3.98
45days
5.95
3.99
3.96
4.07
4.09
4.3
60days
6.5
4.11
4
4.16
4.17
4.4
From the results, we can observe the increase in the peroxide value of the sample
of refined sunflower oil, which is free of any additives, as follows from 3.66 to6.5.
61
While the oil samples, whether containing the herbal extracts or the industrial
antioxidant, the increase in the values of peroxide were few. These results were
represented on the chart.
7
Peroxide value )meq/kg(
6
5
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 20 - Effect of adding BHT and herbal extracts extracted with ethanol 95%
and propylene glycol 95% on peroxide value of sunflower oil
62
The comparison between the different samples shows that the sample that has the
least peroxide value is the oil sample containing the rosemary extract because, rosemary
extract contained the highest concentration of phenolic substances.
Furthermore, the increase in the peroxide values of the oil samples containing the
herb extract is less than the increase in the peroxide value of the oil sample containing the
industrial antioxidant.
Thus, we conclude that herbal extracts have the potential to inhibit the oxidation
process of refined sunflower oil more than the industrial antioxidant.
In the beginning, the acid value was measured for the refined sunflower oil sample,
which did not contain any additives, and the result was 0.114 mg КОН / g.
Then the acid values were measured for different oil samples either containing herb
extracts or an industrial antioxidant every 15 days for two months. The results were as
follow in the table17.
Table 17 – The effects of (BHA), thyme extract, ginger extract, rosemary extract,
added to the refined sunflower oil, on its acid values
Day
The
The sample of
The sample
The sample of
The sample
The
sample of
sunflower oil
of sunflower
sunflower oil
of
sample of
sunflower
with thyme
oil with
with
sunflower
sunflower
oil
extract
rosemary
ginger
oil with
oil with
extract
extract
sage
BHT
without
additions
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.24
0.22
0.24
0.25
0.24
30days
0.47
0.3
0.28
0.33
0.34
0.36
45days
0.65
0.36
0.34
0.44
0.45
0.47
60days
0.87
0.48
0.4
0.53
0.56
0.58
63
Through the results, we note that the acid value of the sample of sunflower oil
without any additions have increased from 0.114 to 0.87.On the other hand, the acid values
of refined sunflower oil samples containing either a mixture of herb extracts or an
industrial antioxidant have not changed significantly. The change in the acid values was
represented on the chart.
1
0.9
Acid value ) mg КОН / g(
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 21 -Effect of adding BHT and herbal extracts extracted with ethanol 95%
and propylene glycol 95% on acid value of sunflower oil
When comparing different samples, the sample that has the least acid value is the
oil sample containing the rosemary extract. This may be because it contains a higher
64
percentage of compounds that are effective as antioxidants. From the results in table17,
we note also that increase in the acid value of the oil sample containing BHT is greater
than the increase in the acid values of all the oil samples containing the herb extracts.
Wherefore, we suggest that the BHT has a less effectiveness in delaying the
occurrence of the process of oxidation of refined sunflower oil than herbal extracts.
3.3.6 Effect of adding a mixture of herb extracts extracted with ethanol
95%and propylene glycol 95% or BHT on the peroxide values and on acid values of
refined sunflower oil samples
In this experiment, a mixture of extracts (thyme- ginger, rosemary- thyme, gingerrosemary, sage- rosemary) was added at concentration 1% to 100 ml of refined sunflower
oil sample and BHT at concentration 180 mg / Kg .In the beginning, the peroxide value
was measured for the refined sunflower oil sample, which did not contain any addition,
and the value was 3.66 meq/kg. After the addition of BHT or a mixture of the herb extracts
to the refined sunflower oil samples, the peroxide values of these samples were
determined every 15 days for 60 days. The results were as follow in thetable18.
Table 18 – Effect of adding synthetic antioxidants or a mixture of herbal extracts
on the peroxide values of refined sunflower oil
Days
The
The
The sample of
The sample of
The sample
The
sample of
sample of
sunflower oil
sunflower oil
of
sample of
with
sunflower
sunflower
sunflower sunflower with rosemary
oil
oil with
and thyme
ginger and
oil with
oil with
without
thyme
extract
rosemary
sage and
BHT
additions
and
extract
rosemary
ginger
extract
extract
Peroxide value (meq/kg)
65
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.74
3.73
3.76
3.78
3.79
30days
5.3
3.85
3.8
3.89
3.91
3.98
45days
5.95
3.93
3.86
4.01
4.03
4.3
60days
6.5
4
3.94
4.11
4.14
4.4
From the results, after 60 days we note that the sample that has the highest peroxide
value the sample of oil free of additives. While we note that the peroxide values of the oil
samples containing either the BHT or a mixture of herbal extracts did not exhibit a
considerable increase during sixty days of storage. The increase in the peroxide values
was represented on the chart.
Peroxide value )meq/kg(
7
6
5
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 22 -Effect of adding BHT and a mixture of herbal extracts extracted with
ethanol 95% and propylene glycol 95% on peroxide value of sunflower oil
66
The results of the experiment showed that the oil sample containing the rosemarythyme extract had the lowest peroxide values. The peroxide value for this sample
(3.94meq/kg) was less than the limit of acceptable values (10 meq/kg), stipulated by the
standard GOST 1129-2013 for sunflower oil, because they contain compounds such as
rosmanol, rosmaridiphenol, and carnosol which are effective as antioxidants. The increase
in the peroxide values of the oil sample containing BHT antioxidant is more than the
increase in the peroxide value of the oil samples containing the mixture of herb extract.
Thus we conclude that a mixture of herbal extracts has the antioxidant efficacy in
stabilizing the oils by delaying the hydroperoxides formation more than the industrial
antioxidant.
At first, the acid value was measured for the refined sunflower oil sample, which
did not contain any additives; it was 0.114.Every 15 days for two months, the acid value
of the oil samples containing mixture of herbal extracts or the synthetic antioxidant was
measured and the results were as follows in the table19.
Table 19 – Effect of adding a mixture of herbal extracts or BHT on the acid values
of refined sunflower oil
Days
The
The
The
The sample of
The sample
The
sample of
sample of sample of
sunflower oil
of sunflower
sample of
sunflower
sunflower sunflower
with
oil with sage
sunflower
oil
oil with
oil with
ginger and
and rosemary
oil with
without
thyme
rosemary
rosemary
extract
BHT
additions
and
and
extract
ginger
thyme
extract
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.19
0.18
0.22
0.2
0.24
30days
0.47
0.28
0.26
0.33
0.34
0.36
67
45days
0.65
0.37
0.31
0.43
0.44
0.47
60days
0.87
0.44
0.36
0.5
0.52
0.58
The results show that after2monthsthe sample that had the highest acid value the
sample of oil without any addition. While the oil samples- whether containing the mixture
of herbal extracts or BHT- the increase in the values of acid were few. The in creasein the
acid values was represented on the chart.
1
Acid value ( mg КОН / g)
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
10
20
30
40
50
60
70
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 23 -Effect of adding BHT and a mixture of herbal extracts extracted with
ethanol 95% and propylene glycol 95% on acid value of sunflower oil
68
Table 20– Regression equations describing the change in acid value for the samples
of sunflower oil within two months
The sample of sunflower oil
The regression equation
Coefficient of
approximation
The sample of sunflower oil without
additions
y = 0.0125x + 0.1049
R² = 0.9972
The sample of sunflower oil with thyme and
ginger extract
y = 0.0071x + 0.1082
R² = 0.9917
The sample of sunflower oil with rosemary
and thyme extract
y = 0.0042x + 0.118
R² = 0.9889
The sample of sunflower oil with ginger and
rosemary extract
y = 0.0066x + 0.1186
R² = 0.9925
The sample of sunflower oil with sage and
rosemary extract
y = 0.0056x + 0.109
R² = 0.9983
The sample of sunflower oil with BHT
y = 0.0078x + 0.1158
R² = 0.9981
y-Acid value (mg КОН / g); x- Storage time (day).
The comparison between the different samples shows that the sample that has the
least acidity value is the oil sample containing the rosemary-thyme extract, because they
contain compounds such as rosmanol, rosmaridiphenol, and carnosol which are effective
as antioxidants
The acid number for this sample (0.36 mg KOH / g of oil) was approaching the
limit of acceptable values (0.4 mg KOH / g of oil), stipulated by the standard GOST 11292013 for sunflower oil.
On the other hand, the acid value of the oil sample containing the synthetic was
greater than the acid values of all samples containing a mixture of herbal extracts.
Therefore, we conclude that synthetic antioxidant has the antioxidant efficacy in
stabilizing the oils by delaying the free fatty acid formation less than a mixture of herbal
extracts.
69
3.3.7 Effect of adding herb extracts extract with propylene glycol 95%
separately or BHT on the peroxide values and on acid values of refined sunflower oil
samples
At the beginning of the experiment, the peroxide value of the refined sunflower oil
sample was measured; the value was 3.66 meq/kg.In this experiment, extracts were added
separately at concentration 1% to 100 ml of refined sunflower oil sample and BHT at
concentration 180 mg / Kg.
After adding herbal extracts and industrial antioxidants to the oil samples separately
and measuring the peroxide values for each sample after 15 days and for two months, the
results were as in the table 21.
Table 21 – The effect of herbal extracts or industrial antioxidants on the acid values
of refined sunflower oil when each was added separately
Days
The
The sample
The sample of
The sample
The sample
The
sample of
of sunflower
sunflower oil
of sunflower
of sunflower
sample of
sunflower
oil with
with rosemary
oil with
oil with sage
sunflower
oil
thyme
extract
ginger
extract
oil with
without
extract
extract
BHT
additions
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
3.66
3.66
15 days
4.6
3.78
3.76
3.78
3.79
3.79
30days
5.3
3.9
3.88
3.93
3.95
3.98
45days
5.95
4.01
3.98
4.09
4.11
4.3
60days
6.5
4.14
4.05
4.17
4.19
4.4
A sample of refined sunflower oil free from herb extracts or BHT increased its
peroxide value more than other samples as it increased from 3.66 to 6.5.
70
On the other hand, the peroxide values of refined sunflower oil samples containing
either of herb extracts or an industrial antioxidant have not changed significantly. The
change in the peroxide values was represented on the chart.
7
6
5
Peroxide value )meq/kg(
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 24 -Effect of adding BHT and herbal extracts extracted with propylene glycol
95% on peroxide value of sunflower oil
71
The sample containing rosemary extracts had the lowest peroxide value. This may
be because it contains a higher percentage of polyphenols than other samples.
It was also found that all the samples of oil containing herbal extracts have lower
peroxide values than the sample containing the synthetic antioxidant.
We can conclude that herbal extracts that have been added to sunflower oil have the
potential to delay the occurrence of oxidation of oil more than the industrial antioxidant.
First, the acid value of the refined sunflower oil was determined without any
additives and the result was 0.114 mg КОН / g.
The acid value of refined sunflower oil samples- whether free of additives or
containing (BHT) or the herbal extracts added separately at concentration 1% to the oilwere determined every 15 days for two months, and the results were shown in table22.
Table 22 – The effect of herbal extracts or BHT antioxidants on the acid values of
refined sunflower oil when each was added separately
Days
The
The sample
The
The sample of
The sample
The
sample of
of sunflower
sample
sunflower oil
of sunflower
sample of
sunflower
oil with
of oil
with
oil with sage
sunflower
oil
thyme
with
ginger
extract
oil with
without
extract
rosemary
extract
additions
BHT
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.25
0.24
0.26
0.27
0.24
30days
0.47
0.32
0.3
0.35
0.36
0.36
45days
0.65
0.38
0.36
0.46
0.47
0.47
60days
0.87
0.5
0.42
0.56
0.57
0.58
Among all oil samples, refined sunflower oil sample without any additives showed
the highest increased in acid value.
72
The acid values of the oil samples did not change much after 60 days except for the
oil sample free from any additives, the results were represented on the following chart.
1
0.9
Acid value ) mg КОН / g(
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme extract
sunflower oil sample with rosemary extract
sunflower oil sample with ginger extract
sunflower oil sample with sage extract
sunflower oil sample with BHT
Figure 25 -Effect of adding BHT and herbal extracts extracted with propylene glycol
95%on acid value of sunflower oil
After comparing the different samples, we note that the sample with the lowest acid
value is the sample containing the rosemary extract. This is due to the high percentage of
73
polyphenols. The results show that after the addition of plant extracts to refined sunflower
oil samples, the acid values of these samples were lower than the value of acid when
adding the BHT antioxidant. Wherefore, we suggest that the BHT antioxidant had the
potential to inhibit the oxidation process of refined sunflower oil less than herbal extracts.
3.3.8 Effect of adding a mixture of herb extracts extract with propylene glycol
95% or BHT on the peroxide values and on acid values of refined sunflower oil
samples
In this experiment, a mixture of extracts (thyme- ginger, rosemary- thyme, gingerrosemary, sage- rosemary) was added at concentration 1% to 100 ml of refined sunflower
oil sample and BHT at concentration 180 mg / Kg. Firstly, the peroxide value of refined
sunflower oil was determined without any addition the value was 3.66 meq/kg. Then the
synthetic antioxidant or a mixture of herbal extracts were added to the oil samples.
Peroxide values were measured for each sample after 15 days and for two months. The
results were as follows in the table 23.
Table 23 – Effect of adding a mixture of herbal extracts or industrial antioxidants
on the peroxide values of refined sunflower oil
Days
The
The
The sample of
The sample of
The sample
The
sample of
sample of
sunflower oil
sunflower oil
of sunflower
sample of
with
oil with sage
sunflower
sunflower sunflower with rosemary
oil
oil with
and thyme
ginger and
and rosemary
oil with
without
thyme
extract
rosemary
extract
BHT
additions
and
3.66
3.66
extract
ginger
extract
Peroxide value (meq/kg)
1day
3.66
3.66
3.66
3.66
74
15 days
4.6
3.75
3.74
3.77
3.78
3.79
30days
5.3
3.86
3.83
3.91
3.93
3.98
45days
5.95
3.95
3.89
4.03
4.07
4.3
60days
6.5
4.02
3.97
4.14
4.17
4.4
Among all oil samples, refined sunflower oil sample without any additives showed
the highest increased in peroxide value. On the other hand, the peroxide values of refined
sunflower oil samples containing either a mixture of herb extracts or BHT antioxidant
have not changed significantly. The change in the peroxide values was represented on the
chart.
Peroxide value )meq/kg(
7
6
5
4
3
2
1
0
1
15
30
45
60
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 26 -Effect of adding BHT and a mixture of herbal extracts extracted with
propylene glycol 95% on peroxide value of sunflower oil
75
The comparison between the different samples shows that the sample containing
the rosemary-thyme extract had the lowest peroxide value. The peroxide value for this
sample (3.97meq/kg) was less than the limit of acceptable values (10 meq/kg), stipulated
by the standard ТС 024/2011 for technical regulations for oil and fat products, because
they contain a higher proportion of polyphenols than other samples.
The results show that all the samples of oil containing a mixture of herbal extracts
have lower peroxide values than the sample containing the synthetic antioxidant.
We can conclude that the mixture of herbal extracts has a greater effectiveness in
delaying the occurrence of the process of oxidation of refined sunflower oil than BHT.
At the beginning of the experiment, the acid value of the refined sunflower oil
sample was measured sample; it was0.114 mg КОН / g. Every 15 days for two months,
the acid value of the oil samples containing the synthetic antioxidant or a mixture of herbal
extracts was measured and the results were as follows in the table 24.
Table 24 – Effect of adding a BHT or a mixture of herbal extracts on the acid values
of refined sunflower oil
Days
The
The
The
The sample of
The sample
The
sample of
sample of
sample of
sunflower oil
of sunflower
sample of
oil with
with
oil with sage
sunflower
sunflower sunflower
oil
oil with
rosemary
ginger and
and
oil with
without
thyme
and thyme
rosemary
rosemary
BHT
additions
and
extract
extract
extract
ginger
extract
Acid value ( mg КОН / g)
1day
0.114
0.114
0.114
0.114
0.114
0.114
15 days
0.31
0.21
0.2
0.2
0.23
0.24
30days
0.47
0.3
0.28
0.34
0.36
0.36
45days
0.65
0.39
0.34
0.44
0.46
0.47
60days
0.87
0.47
0.39
0.52
0.53
0.58
76
The results show that after 2months the sample that had the highest acid value the
sample of oil without any addition. While the oil samples, whether containing the mixture
of herbal extracts or BHT, the increase in the values of acid were few. The results were
represented on the following chart.
1
0.9
Acid value ( mg КОН / g)
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
10
20
30
40
50
60
70
Storage time(day)
sunflower oil (control)
sunflower oil sample with thyme and ginger extract
sunflower oil sample with rosemary and thyme extract
sunflower oil sample with ginger and rosemary extract
sunflower oil sample with sage and rosemary extract
sunflower oil sample with BHT
Figure 27 -Effect of adding BHT and a mixture of herbal extracts extracted with
propylene glycol 95% on acid value of sunflower oil
77
Table 25 – Regression equations describing the change in acid value for the samples
of sunflower oil within two months
The sample of sunflower oil
The regression equation
Coefficient of
approximation
The sample of sunflower oil without
additions
y = 0.0125x + 0.1049
R² = 0.9972
The sample of sunflower oil with thyme and
ginger extract
y = 0.0072x + 0.1223
R² = 0.987
The sample of sunflower oil with rosemary
and thyme extract
y = 0.0071x + 0.1082
R² = 0.9917
The sample of sunflower oil with ginger and
rosemary extract
y = 0.006x + 0.1149
R² = 0.9982
The sample of sunflower oil with sage and
rosemary extract
y = 0.0047x + 0.1238
R² = 0.9848
The sample of sunflower oil with BHT
y = 0.0078x + 0.1158
R² = 0.9981
y-Acid value (mg КОН / g); x- Storage time (day).
The results of the experiment showed that the sample that has the least acid value
is the oil sample container of the rosemary-thyme extract.
The acid number for this sample (0.39 mg KOH / g of oil) was less than the limit
of acceptable values (0.6 mg KOH / g of oil), stipulated by the standard ТР ТС 024/2011
for technical regulations for oil and fat products.
This may be because they contain more polyphenols than other samples.
From the results in the table 24 we note that the acid value of the oil sample, which
is added to it, BHT was higher than the acid values of the oil samples containing a mixture
of the plant extracts.
Wherefore, we conclude that a mixture of herbal extracts has the potential to inhibit
the oxidation process of refined sunflower oil more than BHT.
78
CONCLUSIONS
1.The results show that amount of phenolic compounds for various extracts ranged
from11.12to45.17. The total polyphenol content of the extracts can be ranked in the order
rosemary (45.17)>thyme (36.15) > sage (25.74) > ginger (11.12).The results show that
the rosemary extract contains the highest value of polyphenols.
2. The results show that the DPPH radical scavenging ability of the extracts can be
ranked in the order rosemary (85.15%) >thyme (76.55%) > sage (55.64 %) > ginger
(36.44%). The observed differential scavenging activities of the extracts against the DPPH
system could be due to the presence of different compounds in the extract.
3. In comparing the different samples, we note that the lowest peroxide value was
for the sample of sunflower oil containing the rosemary extract.
On the other hand, the peroxide values of all samples containing herb extracts that
were added separately to these samples were lower than the peroxide values of the oil
sample containing the industrial antioxidant.Thus, we conclude that herbal extracts have
a higher effectiveness in preventing the oxidation of sunflower oil than the industrial
antioxidant.
4. After comparing the different samples, we note that the sample with the lowest
acid value is the sample containing the rosemary extract.
Furthermore, the acid value of the oil sample containing the industrial antioxidant
was greater than the acid values of all samples containing herb extracts that were added
separately to these samples.Thus, we conclude that the industrial antioxidant has a lower
effectiveness in preventing the oxidation of sunflower oil than herbal extracts.
5. The comparison between the different samples shows that the sample containing
the rosemary-thyme extract had the lowest peroxide value.It was also found that all the
samples of oil containing a mixture of herbal extracts have lower peroxide values than the
sample containing the synthetic antioxidant, and as well as below the peroxide values of
79
sunflower oil samples containing extracts that were added separately to these samples .We
can conclude that the mixture of herbal extracts has a greater effectiveness in delaying the
occurrence of the process of oxidation of refined sunflower oil than the synthetic
antioxidant.
6. After comparing the different samples, we note that the sample with the lowest
acid value is the sample containing the rosemary-thyme extract. On the other hand, all the
samples of oil containing a mixture of herbal extracts have lower acid values than the
sample containing the synthetic antioxidant, and as well as below the acid values of
sunflower oil samples containing herbal extracts that were added separately to these
samples.
So, we conclude that the synthetic antioxidant has a less effectiveness in delaying
the occurrence of the process of oxidation of refined sunflower oil than a mixture of herbal
extracts. Therefore, we suggest using a mixture of thyme and rosemary extract as a natural
antioxidant for vegetable oils.
80
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ГОСТ
Р
51487-99.
Масла
растительные
и
жиры
животные.
Методопределенияперекисногочисла.
100. ГОСТ Р 50457-92 Государственный стандарт Российской Федерации.
Жиры и масла животные и растительные. Определение кислотного числа и
кислотности.
101.
ГОСТ 1129-2013 Масло подсолнечное. Технические условия (с
Поправкой).
102.
ТР ТС 024/2011Технический регламент на масложировую продукцию.
91
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