A.

Experiment’s Title
Quantitative Test of Lipids
B. Experiment’s Date
Thursday, November 14th, 2019 at 13.00 WIB
C. Experiment’s Finish
Thursday, November 14th, 2019 at 15.30 WIB
D. Experiment’s Purpose
Determine peroxide number and free fatty acid
E. Basic Theory
1. Lipids
Lipid is any of a diverse group of organic
compounds including fats, oils, hormones, and certain components of
membranes that are grouped together because they do not interact
appreciably with water (Rodrigues and Silva, 2016). One type of lipid,
the triglycerides, is sequestered as fat in adipose cells, which serve as the
energy-storage depot for organisms and also provide thermal insulation
(Eaton and Rogers, 2017). Some lipids such as steroid hormones serve as
chemical messengers between cells, tissues, and organs, and others
communicate signals between biochemical systems within a single cell.
The membranes of cells and organelles (structures within cells) are
microscopically thin structures formed from two layers
of phospholipid molecules (Carra, 2018). Membranes function to separate
individual cells from their environments and to compartmentalize the cell
interior into structures that carry out special functions. So important is this
compartmentalizing function that membranes, and the lipids that form
them, must have been essential to the origin of life itself.
Molecules such as proteins, nucleic acids, and carbohydrates have
an affinity for water and are called hydrophilic. Lipids, however,
are hydrophobic. Some lipids are amphipathic part of their structure is
hydrophilic and another part, usually a larger section, is hydrophobic
(McCance and Huether, 2018). Amphipathic lipids exhibit a unique
behaviour in water: they spontaneously form ordered

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molecular aggregates, with their hydrophilic ends on the outside, in contact
with the water, and their hydrophobic parts on the inside, shielded from the
water. This property is key to their role as the fundamental components of
cellular and organelle membranes (Rogers, 2011).
Although biological lipids are not large
macromolecular polymers (e.g., proteins, nucleic acids, and
polysaccharides), many are formed by the chemical linking of several
small constituent molecules. Many of these molecular building blocks are
similar, or homologous, in structure. The homologies allow lipids to be
classified into a few major groups: fatty acids, fatty
acid derivatives, cholesterol and its derivatives, and lipoproteins. This
article covers the major groups and explains how these molecules function
as energy-storage molecules, chemical messengers, and structural
components of cells.

Figure 1 Lipid
a. Structure
Lipids are composed of the elements carbon, hydrogen and
oxygen, similar to carbohydrates, but contain less water (Insel, 2014).
In fact, lipids are insoluble in water. Fats are an example of a type of
lipid. Lipids play a variety of important functions in the cells. The most
common type of lipids are called triglycerides. Triglycerides are made
up of 3 fatty acid chains attached to a glycerol backbone. Fatty acids
are chains of carbon atoms (between 14 and 22) with the end carbon
possessing a carboxyl group (COOH). The fatty acids in the
triglyceride could be the same, or could have different structures.
Glycerol has three carbons, with 3 OH molecules attached. The glycerol
backbone becomes attached to the three fatty acids through

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a condensation reaction because three molecules of water are formed.
The bond that forms between the fatty acid chain and the glycerol is
called an ester bond (Kent, 2000).

Figure 2 Formation of triglyceride molecule from glycerol and three

fatty acids. Three molecules of water are removed in the conensation
reaction (Kent, 2000)
The structure of the fatty acids influences the structure of the
lipid. In the fatty acid chains, the carbon atoms could have single
bonds between them making the lipid “saturated”. This generates fats
that are usually solid at room temperature. Alternatively, if one or more
of the bonds between the carbon atoms are double bonds, the lipid is
said to be “unsaturated”. If there is one double bond, the triglyceride is
said to be “monounsaturated”, if it has multiple double bonds, it is
“polyunsaturated”. Unsaturated fatty acids are usually liquid at room
temperature and are called oils.
The double bonds in unsaturated fatty acids can exzist in either
a cis or a trans configuration (Lawson, 2013). This describes whether
the hydrogen atom are on the same side (cis) or opposite sides (trans).
A cis double bond generates a bend in the molecule, influencing its
structure and downstream function. Trans fats are rare in nature. Fat

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 molecules with fully saturated tails can pack tightly against one another
because the single bonds result in straight molecules. This tight packing
generates fats that are solid at room temperature, for example, butter.
Unsaturated fatty acids (which in nature usually contain cis double
bonds) have bent tails. This means they are not able to be tightly packed
and results in oils that are liquid at room temperature.

Figure 3 Sructure of saturated, unsaturated, and trans fatty acid

(Lawson, 2013)
b. Characteristics
1) Lipids may be either liquids or non-crystalline solids at room
temperature.
2) Pure fats and oils are colourless, odorless, and tasteless.
3) Also known as calorie density, energy density is the total amount of
calories per specific weight of food. Lipids have high energy
density, which means the amount of energy they release per unit of
mass is high. 1 gram of a lipid contains 9 calories per gram, which
is about 2.25 times greater than proteins and carbohydrates. The
inability of lipids to dissolve in water and hence bond with it means
that no excess weight is added to their existing mass without any
incremental increase in calorie density.

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 4) With the exception of phospholipids, which partially dissolve in
water, all lipids are generally insoluble in water. This is due to the
presence of long-chain fatty acids in their structure. The cell
membranes of your body are composed of phospholipids. They help
create a semi-permeable barrier between the cells and the external
environment. If water-soluble compounds were used as components
of cell membranes, they wouldn’t be as durable since they would
dissolve in water.
5) Solid triglycerols (Fats) have high proportions of saturated fatty
acids.
6) Liquid triglycerols (Oils) have high proportions of unsaturated fatty
acids.
c. Function
In the human body, triglycerides are mostly stored in fat cells,
called adipocytes, which form adipose tissue. Adipose tissue is
primarily used as an energy store, but also helps
to protect and insulate the body. Lipids have a variety of functions in
the cell.
a. Energy storage
Triglyceride breakdown yields more energy than the breakdown of
carbohydrates because the carbons are all bonded to hydrogens (and
they, therefore, have a higher proportion of hydrogens relative to
oxygens) (Gurr, 2012). This means they are electron-rich and can
contribute to the production of acetyl-CoA, which is an important
co-enzyme in aerobic respiration.
b. Biological membranes
As previously discussed, cell membranes are principally composed
of a phospholipid bilayer. Phospholipids are another type of lipid,
created when a phosphate group replaces one of the three fatty acid
chains. Phospholipids have a hydrophobic part and a hydrophilic
part. The fatty acid chains remain hydrophobic, forming the tail of
the molecule, but the addition of the phosphate group to the head

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 makes this part of the molecule hydrophilic, meaning a bilayer
forms (Gurr, 2012).
c. Hormone production
Many hormones are lipid-derived, and they usually belong to a class
of hormones called steroid hormones. These hormones are usually
derived from cholesterol and are often structurally similar to
cholesterol. Steroid hormones are important signalling molecules
that can enter the cell directly through the cell membrane and
influence gene expression and signalling pathways. Examples
include cortisol and testosterone.
2. Palm Oil
Palm oil originated in western Guinea and is now cultivated in other
tropical countries. Mostly it is obtained from the fruit of the African palm.
Palm oil is orange-red in color due to its beta-carotene content (Marcus,
2013). It is used in cooking oil, margarine, and rich-tasting processed
foods. If the oil is boiled, the carotenoid content may be destroyed, and the
oil may become colorless. One tablespoon of palm oil contains 119
calories, 13.5 grams of total fat, 6.7 grams of saturated fat, 5 grams of
monounsaturated fats, and 1.3 grams of polyunsaturated fats (Marcus,
2013). Palm oil has a good stability at the high temperatures used in frying
(usually 175–185 °C), because of its content of natural antioxidants, the
absence of highly unsaturated fatty acids, and the moderate content
of linoleic acid. Consequently, palm oil or palm olein is widely used
domestically and in industry.
Most of the fatty acids which contained in palm oil are saturated
fatty acids namely palmitic acid (Rival and Levang, 2014). Saturated fatty
acids only have a single bond between the constituent carbon atoms,
whereas unsaturated fatty acids have at least one double bond between the
constituent carbon atoms. Saturated fatty acids are more stable than
unsaturated fatty acids. Double bonds in unsaturated fatty acids easily react
with oxygen (easily oxidized). The existence of a double bond in

Page | 6
 unsaturated fatty acids makes it has two forms: cis which is unstable and
trans that is stable (Gunstone, 2012).
The physical and chemical properties of palm oil include color, odor,
flavor, solubility, melting point and polymorphism, boiling point, flash
point and fire point, iod number, and saponification number is depending
on the purity and quality of palm oil. Some physical and chemical
properties of palm oil generally can be seen in the table below.

Figure 4 The composition content of palm oil

Figure 5 The composition of fatty acid in palm oil

The color of the oil is determined by the presence of pigment
remaining after the bleaching process. Smell and flavor in oil are naturally
present, also occur due to the presence of short-chain fatty acids due to oil
damage (Gunstone, 2012).
3. Fatty Acid

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Fatty acid is an important component of lipids (fat-soluble components of
living cells) in plants, animals, and microorganisms (Rodrigues and Silva,
2016). Generally, a fatty acid consists of a straight chain of an even number
of carbon atoms, with hydrogen atoms along the length of the chain and at
one end of the chain and a carboxyl group (―COOH) at the other end. It is
that carboxyl group that makes it an acid (carboxylic acid). If the carbon-
to-carbon bonds are all single, the acid is saturated; if any of the bonds is
double or triple, the acid is unsaturated and is more reactive (Deeker, et al,
2010). A few fatty acids have branched chains; others contain ring
structures (e.g., prostaglandins). Fatty acids are not found in a free state in
nature; commonly they exist in combination with glycerol (an alcohol) in
the form of triglyceride.
Among the most widely distributed fatty acids are the 16- and 18-carbon
fatty acids, otherwise known as palmitic acid and stearic acid, respectively.
Both palmitic and stearic acids occur in the lipids of the majority of
organisms. In animals palmitic acid makes up as much as 30 percent of
body fat. It accounts for anywhere from 5 to 50 percent of lipids in
vegetable fats, being especially abundant in palm oil. Stearic acid is
abundant in some vegetable oils.
a. Saturated Fatty Acid
The saturated fatty acids are derived from both animal fats and plant
oils. Rich sources of dietary saturated fatty acids include butter fat, meat
fat, and tropical oils (palm oil, coconut oil, and palm kernel oil).
Saturated fatty acids are straight-chain organic acids with an even
number of carbon atoms. The simplest fatty acids are unbranched,
linear chains of CH2 groups linked by carbon-carbon single bonds with
one terminal carboxylic acid group. The term saturated indicates that
the maximum possible number of hydrogen atoms are bonded to each
carbon in the molecule. Many saturated fatty acids have a trivial or
common name as well as a chemically descriptive systematic name.
The systematic names are based on numbering the carbon atoms,
beginning with the acidic carbon. The table gives the names and typical

Page | 8
 biological sources of the most common saturated fatty acids. Although
the chains are usually between 12 and 24 carbons long, several shorter-
chain fatty acids are biochemically important. For instance, butyric acid
(C4) and caproic acid (C6) are lipids found in milk. Palm kernel oil, an
important dietary source of fat in certain areas of the world, is rich in
fatty acids that contain 8 and 10 carbons (C8 and C10).
b. Unsaturated Fatty Acid
Unsaturated fatty acids have one or more carbon-carbon double bonds.
The term unsaturated indicates that fewer than the maximum possible
number of hydrogen atoms are bonded to each carbon in the molecule.
The number of double bonds is indicated by the generic name
monounsaturated for molecules with one double bond or
polyunsaturated for molecules with two or more double bonds. Oleic
acid is an example of a monounsaturated fatty acid. Common
representative monounsaturated fatty acids together with their names
and typical sources are listed in the table. The prefix cis-9 in the
systematic name of palmitoleic acid denotes that the position of the
double bond is between carbons 9 and 10. Two possible conformations,
cis and trans, can be taken by the two CH2 groups immediately adjacent
to the double-bonded carbons. In the cis configuration, the one
occurring in all biological unsaturated fatty acids, the two adjacent
carbons lie on the same side of the double-bonded carbons. In the trans
configuration, the two adjacent carbons lie on opposite sides of the
double-bonded carbons.
4. Peroxide Number
The determination of the peroxide value is the traditional and most used
parameter for measuring the primary products of oxidative degradation.
From this value, the propagation step of the free radical chain mechanism
and the accumulation of hydroperoxides can be followed (Decker, et al,
2010). However, it is not possible to use the peroxide value alone to judge
the quality of edible oils, because hydroperoxides decompose during
storage. This decomposition can take place faster than the formation of new

Page | 9
 hydroperoxides, depending on certain storage conditions such as
temperature, light or metal traces. Although the oil has already been
damaged by oxidation, and higher levels of degradation products have
already formed, the speed of hydroperoxide decomposition can result in
falsely low levels of these compounds. In this instance, the peroxide value
tells us nothing about the real quality of the product. In order to avoid
misinterpreting peroxide values, it is necessary to know the history of the
sample. However, the peroxide value is a suitable parameter for measuring
the deterioration of quality over time. After the induction period, during
which the peroxide value increases slowly, a steep increase indicates that
the oil has been gone bad.
In general, the aim of oil production should be to produce oils with peroxide
values as low as possible, without the formation of secondary reaction
products. A higher peroxide value at the beginning of the storage period
has a negative effect on the storage stability of the oil. For refined oils,
producers should aim for a peroxide value below 1, better 0.5 meq O2/kg
oil, while the peroxide value for virgin oils can be higher, up to 3 meq O2/kg
oil. Based on SNI 01-3741-2002 the good quality of oil has peroxide value
maximal 10 meq/kg.

Figure 6 Forming peroxide mechanism

5. Determining the Peroxide Number using Permanganometry Titration
Permanganometry is a redox titration that uses KMnO4 as a titrant.
Potassium permanganate is a strong oxidizing agent. KMnO4 can be
obtained easily, is inexpensive and requires no indicators except for very
dilute solutions. Permanganate reacts quickly with many reducing agents

Page | 10
 based on the reaction, but there is a need to warm up or use a catalyst to
speed up the reaction.
The deep purple color of KMnO4 is useful in making the
permanganate itself an indicator of the titration. One drop of excess can
produce bright colors even in large volumes of solution. Potassium
permanganate is a strong oxidizer in acidic solutions and does not require
indicators. In such an environment the permanganate ion is reduced to four-
dimensional manganese
6. Free Fatty Acid
Free fatty acids are presented in crude oils but they are removed during
refining process. Free fatty acids are more susceptible to autoxidation than
esterified fatty acids. Thus, free fatty acids act as pro-oxidants in edible oil.
These compounds have a hydrophilic and a hydrophobic group in their
structure. The carbonyl group is the hydrophilic group and the hydrocarbon
chain is the hydrophobic group (Erickson, 1990). The carbonyl group of
these compounds are preferably concentrated on the surface of edible oil,
decreasing the surface tension and increasing the diffusion rate of oxygen
from the headspace into the oil, so accelerating oil oxidation. Picture below
shown how the free fatty acid formed from triglyceride.

Regarding mono- and diacylglycerol, they are present in crude oils. These
compounds also act as pro-oxidants according to their chemical structure.
The hydrophilic groups in these compounds are the hydroxyl groups. Their
effect is similar than the effect of free fatty acids facilitating the diffusion
of the oxygen. For that reason, they should be removed during the refining

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 process. Based on SNI 01-3741-2002 the good quality of oil has FFA value
maximal 0.3%.

Figure 7 Syarat mutu minyak goreng

7. Determining the Free Fatty Acid using Acidimetry Titration
The determination of the strength of a solution of an acid by titration with
a standard solution of a base is called acidimetry. The completion of the
reaction between an acid an an alkali is termed neutralization and it
proceeds with the formation of a salt and water (Akhluwalia, Dhingra and
Gulati, 2005). The detection of the end point of the titration is assisted by
the addition of an indicator. In this experiment, to determine the free fatty
acid in the oil can use the phenolphthalein indicator. The indicator
employed in these titration is required to indicate the equivalence point
rather than the true neutral point. At the point of equivalence, the pH of the
solution could be equal to, greater than or less than 7, depending on the
relative strengths of the acid an alkali.
Indicators
The indicators employed in acidimetry are either weak organic acids or
weak organic bases. Their degree of dissociation is greatly affected by any
alteration of the hydrogen ion concentration of the solution. The choice of
a suitable indicator for a particular titration depends upon the nature of the
acid and base used in that titration. A suitable indicator is that which

Page | 12
 changes color at the pH of the equivalence point. Picture below shown the
pH range of an indicator.

Figure 8 pH range of several indicators (Akhluwalia, Dhingra and Gulati,

2005)
Usually an indicator such as phenolphthalein is used for titration involving
a strong alkali like natrium hydroxide. Picture below shows the structure of
phenolphthalein when it reacted with alkali or acid.

Figure 9 Phenolphtalein mechanism reaction with acid or base

(Akhluwalia, Dhingra and Gulati, 2005)

F. Tools and Materials

1. Tools
a. Beaker glass 100 ml 6 pieces
b. Erlenmeyer 100 mL 6 pieces
c. Measuring glass 10 mL 1 piece
d. Measuring glass 50 mL 1 piece

Page | 13
 e. Drop pipette 6 pieces
f. Vial glass 11 pieces
g. Burette (dark) 1 piece
h. Burette 1 piece
i. Statif and clamp 1 set
j. Analytical balance 1 unit
2. Materials
a. Palm oil (new) ± 30 mL
b. Palm oil (waste) ± 30 mL
c. Alcohol 96% 60 mL
d. Aquades 180 mL
e. H2SO4 4N 60 mL
f. KMnO4 0.1 N 5 mL
g. Phenolphtalein indicator 30 drops
h. NaOH 0.1 N Sufficiently
G. Lanes Work
1. Determine Peroxide Number

5 mL oil/fat

11. Entered into Erlenmeyer 100 mL

12. Added 45 mL aquades
13. Added 15 mL H2SO4 4N solution
14. Titrated with KMnO4 0,1 N until pink
solution
Volume KMnO4 0,1 N

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2. Determine Free Fatty Acids
a. Blank solution
6 grams aquades

5. Entered into Erlenmeyer 100 mL

6. Added 10 mL alcohol 96%
7. Added 5 drops PP indicator
8. Titrated with NaOH 0,1 N until pink solution (not
disappear for 30 seconds)
Volume NaOH 0,1 N

b. Sample

6 grams aquades

1. Entered into Erlenmeyer 100 mL

2. Added 10 mL alcohol 96%
3. Added 5 drops PP indicator
4. Titrated with NaOH 0,1 N until pink solution (not
disappear for 30 seconds)
5. Repeated 3x
Volume NaOH 0,1 N

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 H. Observation Result
No Observation Result
Lanes Work Reaction/Assumption Conclusion
Exp Before After
1. 1. Determine Peroxide Number  Aquades : 1. Palm oil (new)  2 KMnO4 + 5H2O2  The peroxide
5 mL oil/fat Colorless  Palm Oil + 45 ml + 3 H2SO4→ number of
solution Aquades : 2 Layers 2MnSO4 + K2SO4 + new oil is
1. Entered into Erlenmeyer
100 mL  H2SO4 4N : (Up : Oil yellow 8H2O + 5O2 0,035 meg/Kg
2. Added 45 mL aquades Colorless solution and Down  MnO4- + 5e- + 8H+  The peroxide
3. Added 15 mL H2SO4 4N
solution solution : Aquades) → Mn2+ + 4H2O number of
4. Titrated with KMnO4 0,1 N  + 15 ml H2SO4 4N
 KMnO4 0,1 N  H2O2 → 2H+ + O2 wasted oil is
until pink solution
5. Repeated 2x : Purple : (Up : Yellow Oil + 2e- 0,069 meg/Kg
Solution and Down :  2MnO4- + H2O2 +
Volume KMnO4 0,1 N
 Palm oil (new) Colorless solution) 6H+ → 2Mn2+ + 5 This result value
: Yellow  After Titrated with O2 + 8H2O isn’t more than
solution KMnO4 : Pink theory, so it is
 Waste palm solution (Upside) Based on the theory, indicate that the
oil : Dark and yellow Peroxide number of quality of palm oil
brown solution solution palm oil is 10 meg/Kg is good enought.
(Downside) (SNI 01-3741-2002)
 V1 : 0,1 mL
 V2 : 0,1 mL

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Palm Oil (Waste)
 Palm Oil + 45 ml
Aquades : 2 Layers
(Up : Oil brown
solution and Down
: Aquades)
 + 15 ml H2SO4 4N
: (Up : Dark brown
oil and Down :
Colorless solution)
 After Titrated with
KMnO4 : Pink
solution (Upside)
and dark brown
solution
(Downside)
 V1 : 0,2 mL
 V2 : 0,2 mL

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2. 2. Determine Free Fatty Acids  .6 gram of  Aquades +  CH3(CH2)14COOH  The FFA of
c. Blank solution aquades : Alcohol : (aq) + NaOH(aq) New Oil is
Colorless Colorless → 0,057 %
6 grams
aquades liquid solution CH3(CH2)14COONa  The FFA of
1. Entered into Erlenmeyer
 Alcohol 96 %  + PP : Colorless (aq) + H2O (l) Wates Oil is
100 mL
: Colorless  After titrated 0,078 %
2. Added 10 mL alcohol 96%
solution with NaOH : Based on Theory, FFA The result value
3. Added 5 drops PP indicator
 3 Drops of PP Pink solution Palm oil = 0,3 % isn’t more than 0,3
4. Titrated with NaOH 0,1 N
Indicator :  V NaOH I : 0,1 (SNI 01-3741-2002) % (Based on SNI)
until pink solution (not
Colorless mL it is indicate that
disappear for 30 seconds)
solution the quality of palm
Volume NaOH
0,1 N  NaOH : oil is good
Colorless enought.
solution

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d. Sample  6 gram of Palm Oil (New)
6 grams aquades palm oil (new)  Palm Oil +

1. Entered into Erlenmeyer : Colorless Alcohol : 2

100 mL liquid Layers (Up : Oil

2. Added 10 mL alcohol  6 gram of yellow solution

96% palm oil and down :

3. Added 5 drops PP (waste) : colorless

indicator Colorless solution)

4. Titrated with NaOH 0,1 liquid  + PP Indicator: 2

N until pink solution  Alcohol 96 % Layers (Up : Oil

(not disappear for 30 : Colorless yellow solution

seconds) solution and down :

Volume NaOH 0,1 N  3 Drops of PP colorless
Indicator : solution)
Colorless  After titrated
solution with NaOH :
 NaOH : 2Layers (Up :
Colorless Pink solution and
solution down : yellow
solution)
V NaOH I : 0,2 mL

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V NaOH II : 0,2 mL
V NaOH III : 0,3 mL

Palm Oil (Waste)

 Palm Oil +
Alcohol : 2
Layers (Up : Oil
dark brown
solution and
down : colorless
solution)
 + PP Indicator: 2
Layers (Up : Oil
dark brown
solution and
down : colorless
solution)
 After titrated
with NaOH : 2
Layers (Up :
Pink solution and

Page | 20
 down : dark
brown solution)
V NaOH I : 0,3 mL
V NaOH II : 0,25 mL
V NaOH III : 0,3 mL

Page | 21
Page | 22
I. Analysis and Explanation
Lipid is any of a diverse group of organic
compounds including fats, oils, hormones, and certain components of
membranes that are grouped together because they do not interact appreciably
with water (Rodrigues and Silva, 2016). The most common type of lipids are
called triglycerides. Triglycerides are made up of 3 fatty acid chains attached
to a glycerol backbone. Fatty acids are chains of carbon atoms (between 14 and
22) with the end carbon possessing a carboxyl group (COOH). In the fatty acid
chains, the carbon atoms could have single bonds between them making the
lipid “saturated”. If one or more of the bonds between the carbon atoms
are double bonds, the lipid is said to be “unsaturated”. Most of the fatty acids
which contained in palm oil are saturated fatty acids namely palmitic acid
(Rival and Levang, 2014). The quality of an oil can be seen by the value of
peroxide number and free fatty acid number.
In this experiment entitled quantitative test of lipids aims to determine
the peroxide number and free fatty acid number of palm oil sample. The quality
of an oil can be seen by the value of peroxide number and free fatty acid. The
peroxide number is an index of the amount of fat or oil that has been oxidized
(Lawson, 2012). However, it is not possible to use the peroxide value alone to
judge the quality of edible oils, because hydroperoxides decompose during
storage. This decomposition can take place faster than the formation of new
hydroperoxides, depending on certain storage conditions such as temperature,
light or metal traces. While the free fatty acid is the product of hydrolysis
process of triglyceride acid. The level of FFA depends on time, temperature
and moisture content because the oild and fats are exposed tovarious
environments such as storage, processing, heating or frying. The content of free
fatty acid arises, indicating a decrease in quality or damage stage of an oil.
In this experiment, we will use the two different oil namely palm oil
new which hasn’t been used for cooking and the palm oil which has been used
for cooking. Both come from oil with the same brand. There will be two step
in this experiment, namely determining the peroxide number and determining
the free fatty acid.

Page | 23
1. Determine the Peroxide Number
The peroxide value is defined as the amount of peroxide oxygen per
1 kilogram of fat or oil (Insel, 2014). The peroxide value (PV) is a very
important characteristic of lipid quality. It is an index of the amount of fat
or oil that has been oxidized. The assessment of hydro peroxides provides
an estimate of the overall oxidation status for lipids and lipid-containing
foods especially in the primary phase of oxidation, generally known as the
induction period. From this value, the propagation step of the free radical
chain mechanism and the accumulation of hydro peroxides can be
followed (Decker, etal, 2010). The higher the peroxide number the lower
quality of an oil. The high peroxide value can be affect by several factors
such as temperature, air, metal catalyst, or any other factors that can
accelerate the oxidation process of an oil.
The oil that we use in this experiment is palm oil. One tablespoon of
palm oil contains 119 calories, 13.5 grams of total fat, 6.7 grams of
saturated fat, 5 grams of monounsaturated fats, and 1.3 grams of
polyunsaturated fats (Marcus, 2013). Palm oil has a good stability at the
high temperatures used in frying (usually 175–185 °C), because of its
content of natural antioxidants, the absence of highly unsaturated fatty
acids, and the moderate content of linoleic acid. Most of the fatty acids
which contained in palm oil are saturated fatty acids namely palmitic acid
(Rival and Levang, 2014). The oil sample that we used in this experiment
for new palm oil and after used palm oil is oil which has trademark as
“Bimoli”.
The basic principle to determine the peroxide value is redox titration
using KMnO4 as a standard, so it also called permanganometric titration.
Potassium permanganate is a strong oxidizing agent. KMnO4 can be
obtained easily, is inexpensive and requires no indicators except for very
dilute solutions. In this reaction, MnO4 ions act as oxidizing agents. MnO4
ions will turn into Mn2+ ions in an acidic atmosphere. Permanganate reacts
quickly with many reducing agents based on the reaction. The deep purple
color of KMnO4 is useful in making the permanganate itself an indicator

Page | 24
of the titration. One drop of excess can produce bright colors even in large
volumes of solution.
The first step of this experiment is preparing 4 different erlenmeyer,
2 of them filled with 5 mL new palm oil, and the other two of them filled
with 5 mL after used palm oil. Then after that added 45 mL aquades into
each erlenmeyer. The function of adding aquades is to make the
observation of the color change in the titration process will be more easy
to observe because the addition of aquades will increase the volume of the
solution, so when the titration process was occur the end point of the
titration will be easier to observed. Other than that, the function of adding
aquades is to solve the hydrogen peroxide, in other words it can be use as
a solvent. Hydrogen peroxide has two equivalent central atoms with two
unshared pairs and two single bonds. This creates an angled bond, creating
the bent shape of the molecule. Also, because the atoms do not all lie in
the same plane, the dipoles do not cancel each other out. So, hydrogen
peroxide is a polar molecule (Moore, 2013). While water was also polar
molecule, it is polar because of the bent shape of the molecule. The shape
means that most of the negative charge of oxygen is on the side of the
molecule and the positive charge of the hydrogen atom is on the other side
of the molecule. This is an example of a polar covalent chemical bond.
And through the principle of like-dissolve-like, hydrogen peroxide can be
dissolve in the water because it is also polar compound.

Figure 10 Structure of H2O2 Figure 11 Structure of H2O

Page | 25
Here is formation peroxide mechanism:

After that we added 15 mL H2SO4 4 N solution for each erlenmeyer. The

function of adding the strong acid like H2SO4 is to make an acid condition
for the solution so that the redo reaction can be occur. This
permanganometry titration must be carried out in an acidic atmosphere,
because in an acidic atmosphere the ion can be oxidized, whereas in an
alkaline or neutral atmosphere the result will form a precipitate. Sulfuric
Acid is used because the potential oxidation is affected by the
concentration of hydrogen ions but the concentration of Mn2+ doesn’t
influenced the potential redox, the concentration of Mn2+ was able to
reduce permanganate ion by forming Mn3+ and MnO2. In other words,
when H2SO4 replaced with base, the permanganate ion will be reduced as
follows:
MnO4- + e- → MnO42- Eo = 0.5 volt (Svehla, 1995)
If the H2SO4 replaced with neutral solution, the permanganate ion will be
reduced to become mangan dioxide as follows :
MnO4- + 4H+ + 3e- → MnO2 + 2H2O Eo = 1.70 volt (Svehla, 1995)
If the H2SO4 replaced with weak acid, the oxidation number of MnO4- will
change from 7 to become 4, it means that the MnO4- will be reduced to
become MnO2 which is formed brown precipitation. So in this titration,
permanganate should be reduce to become Mn2+ so that, if the reduction
potential are increase permanganate will hard to reduced by reducing agent
or in this case is H2O2 that has Eo = 1.78 volt. The formed of mangan

Page | 26
dioxide from reduction permanganate in the weak acid condition will
formed precipitation that will disturb the titration process because it can
cause the trouble to observe the end point of the titration.
Then we titrated each erlenmeyer with standart solution KMnO4 0,1 N
until the color of the solution change to become purple. Then we repeated
the titration process twice for each erlenmeyer. In this titration process
doesn’t need any indicator because the MnO4- ion has purple color which
can be used as indicator it self so it can called as autoindicator. The deep
purple color of KMnO4 is useful in making the permanganate itself an
indicator of the titration. One drop of excess can produce bright colors
even in large volumes of solution. Potassium permanganate is a strong
oxidizer in acidic solutions and does not require indicators. The end point
of the titration identify with the change of the solution to become purple,
this change due to because the formation of ion Mn2+ which produced form
redox reaction between hydrogen peroxide and permanganate. The
reaction is :
2KMnO4 + 5H2O2 + 3 H2SO4→ 2MnSO4 + K2SO4 + 8H2O + 5O2
MnO4- + 5e- + 8H+ → Mn2+ + 4H2O
H2O2 → 2H+ + O2 + 2e-
2MnO4- + H2O2 + 6H+ → 2Mn2+ + 5 O2 + 8H2O
In the reaction above, the permanganate MnO4- which has oxidation
number +7 reduced by hydrogen peroxide to become Mn2+ which has
oxidation number +2, so it means that the permanganate act as oxidator
and thehydrogen peroxide is a redactor. Due to the potential reduction of
hydrogen peroxide (1.78 volt) which is bigger that MnO4- (1.51 volt) it
become the reason why permanganate reduced and why the hydrogen
peroxide oxidized. From the titration process, we can calculate the
peroxide number with this following formula :
VKMnO4 x NKMnO4 x fp x 17
% H2O2 = x 100 %
5000

Page | 27
Then we get the data as follows :
Oil Average KMnO4 Peroxide Number
volume (mL) (meq/kg)
New 0.1 0.035
After used-oil 0.2 0.069
Based on the SNI 01-3741-2002 the good quality of oil has peroxide value
maximal 10 meg/Kg. From the data we know that the peroxide value from
both oil namely new palm oil and after used palm oil is under the maximum
limit. So, it means that both of the oil is still in good quality and it is still
save to be used to cooking. Although the after-used oil has been used for
several time, the peroxide number is still under the maximum limit, it can
cause by several factors such as the after-used oil might not undergoing
long heating process. When the oil that has been used not too long to left
to contact with open air,the oxidation of fatty acid might not go well, or
maybe the fatty acid isn’t much oxidized. Peroxide number produced from
oxidation process of fatty acid with air or oxygen, when the oil isn’t much
oxidized the peroxide number won’t be significantly different from the
new palm oil. The other reason is, the palm oil has natural antioxidant. In
order to shield the palm oil from oxidative damage during extended storage
and transportation, stabilization of the oil with antioxidants is undoubtedly
indispensable to preserve the good oxidative status of the palm oil. There
are two different antioxidant in palm oil. Those antioxidant source may
comes naturally from the palm oil or synthetic antioxidant which added in
the oil. Active molecules of antioxidants thwart peroxide formation by
binding to oxygen. Antioxidants from oils for food are usually a form of
phenolic. The mechanism of action of antioxidants has two functions. The
first function is the main function of antioxidants, namely as a hydrogen
atom. Antioxidants (AH) which have these main functions are often
referred to as primary antioxidants. These compounds can give hydrogen
atoms quickly to lipid radicals (R *, ROO *) or convert them to a more
stable form, while antioxidant radical derivatives (A *) have a more stable
state than lipid radicals. The second function is a secondary function of

Page | 28
 antioxidants, that is, slowing the rate of auto-oxidation by various
mechanisms beyond the mechanism of breaking the auto-oxidation chain
by converting lipid radicals to a more stable form (Gordon, 1990).

2. Determination of Free Fatty Acids (FFA)

Free fatty acids are fatty acids that are free acids that are not bound
as triglycerides. Free fatty acids produced by the process of hydrolysis and
oxidation usually combine with neutral fat. The results of the hydrolysis
reaction of palm oil are glycerol and free fatty acids. This reaction is
accelerated by factors such as heat, water, acidity and catalysts (enzymes).
The longer the reaction lasts, the more free fatty acid levels are formed.
The oil that we use in this experiment is palm oil. One tablespoon of
palm oil contains 119 calories, 13.5 grams of total fat, 6.7 grams of
saturated fat, 5 grams of monounsaturated fats, and 1.3 grams of
polyunsaturated fats (Marcus, 2013). Palm oil has a good stability at the
high temperatures used in frying (usually 175–185 °C), because of its
content of natural antioxidants, the absence of highly unsaturated fatty
acids, and the moderate content of linoleic acid. Most of the fatty acids
which contained in palm oil are saturated fatty acids namely palmitic acid
(Rival and Levang, 2014). The oil sample that we used in this experiment
for new palm oil and after used palm oil is oil which has trademark as
“Bimoli”.
The basic principle to determine the free fatty acid in the palm oil is
using the acidimetry titration which is usually called as neutralization
titration. Acidimetry is titration method to determine the strength of a
solution of an acid by titration with a standard solution of a base. The
detection of the end point of the titration is assisted by the addition of an
indicator. In this experiment, to determine the free fatty acid in the oil can
use the phenolphthalein indicator. This method choosen for this
experiment because free fatty acid has an acid properties from carboxyl
groups, this acid can be neutralize with base like NaOH. The higher the
free fatty acid in the oil the more volume of NaOH used in titration.

Page | 29
 In this experiment we use blank solution as a correction and
comparison factors towards sample. Blank solution is solution which has
the same treatment with sample tested and has the same content with
sample except the analyte compenent (Laksi, 2000). In this experiment,
blank has the same treatment as sample,it doesn’t contain oil but water. In
this experiment blank solution use to know the exact equivalence point.
Because, the titration is using indicator, and every indicator has pH range.
Other than that, in the sample also contain water, so the function of blank
in this experiment is to know the exact volume of NaOH which reacted
with the oil.
The first step of this experiment is weighing 6 grams of aquades,
three sample of new oil which is each sample contain 6 grams of new oil,
and three sample of after-used oil which is each sample contain 6 grams of
after-used oil using analytical balance. After that, we prepare some
different erlenmeyer which contain each of the sample. The next step is
added 10 mL ethanol 96% into each erlenmeyer.The function of adding
the ethanol 96% is to dissolve the fatty acid which containin the sample.
Ethanol is semipolar because ethanol has OH group and R group, and the
fatty acid structure has polar and non polar group, the longer the carbon
group of fatty acid make the fatty acid tend to be non polar. Because of
this, ethanol use to dissolve the fatty acid completely. Most of the fatty
acids which contained in palm oil are saturated fatty acids namely palmitic
acid. Ethanol is a semi-polar solvent, which means it can dissolve polar
and non-polar compounds. That is why ethanol can also be mixed with
water. The polarity of ethanol is caused by the presence of the -OH group
which is polar, while the ethyl group (CH3CH2-) is a non-polar group.
With a short carbon chain causing ethanol will be semi-polar. Here is the
structure of ethanol, the black area is ethyl group which is non-polar and
the red area is carboxyl group which is polar.

Page | 30
 Non polar

Polar

The next step is added 3 drops of phenolphthalein indicator for each

erlenmeyer. Indicators use to determine the end point of the reaction which
also indicate the equivalence point of the titration where the amount of free
fatty acid has been completely reacted with NaOH. Phenolphtalein
indicator has range pH 8.3-10. Phenolphthalein is an organic compound
(C20H14O4) used as an acid-base indicator. The compound is colorless in
acidic solution and pinkish in basic solution. After that we titrated the
solution using NaOH as a standard until the color of the solution become
pink. The color change in phenolpthalein is due to a change in structure of
the molecule. In acid, the molecule is in its HIn form. This structure
contains a central 5 membered ring which is somewhat strained. In base
the In- structure opens up and becomes flatter. This allows the electrons
more freedom, and the molecule's absorption spectrum now transmits red
light. Thus there is a change in color caused by the change in molecular
shape. Here is the formation of phenolphthalein indicator:

Page | 31
Here is the acid-base reaction during titration:
CH3(CH2)14COOH (aq) + NaOH(aq) → CH3(CH2)14COONa (aq) + H2O
(l)
From the titration we can calculate the FFA content with the volume NaOH
data using this formula:
(𝑉𝑡𝑖𝑡𝑟𝑎𝑡𝑖𝑜𝑛 −𝑉𝑏𝑙𝑎𝑛𝑘 )𝑥 𝑁 𝑁𝑎𝑂𝐻 𝑥 𝐵𝑀 𝑝𝑎𝑙𝑚𝑎𝑡𝑖𝑐 𝑎𝑐𝑖𝑑
%FFA = 𝑥 100 %
𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 𝑥 1000

The, we get the data as follows:

Oil NaOH volume (mL) Average FFA (%)

New V1 = 0.2 0.057
V2 = 0.2
V3 = 0.3
After used-oil V1 = 0.3 0.078
V2 = 0.25
V3 = 0.3

Free fatty acid is another indicator that can express the quality of an oil.
This is because the free fatty acid number can be used to determine the
amount of free fatty acid in the sample. The greater free fatty acid can be
caused by hydrolysis process of triglyceride Based on the SNI 01-3741-
2002 the good quality of oil has free fatty acid content maximal 0.3%.
From the data we know that the free fatty acid content from both oil namely
new palm oil and after used palm oil is under the maximum limit. It means
that the oil is good enough to consume. Although the after-used oil has
been used for several time, the free fatty acid content is still under the

Page | 32
 maximum limit, it can cause by several factors such as the after-used oil
might not undergoing long heating process.
J. Conclusion
1. The peroxide number of new oil is 0,035 meg/Kg while the peroxide
number of wasted oil is 0,069 meg/Kg this result value isn’t more than
theory, so it is indicate that the quality of palm oil is good enough.
2. The FFA of New Oil is 0,057 % while the FFA of Wates Oil is 0,078 %The
result value isn’t more than 0,3 % (Based on SNI) it is indicate that the
quality of palm oil is good enough.
K. Answer of Questions
1. Write down all the reactions that accompanied the fatty acid test in this
experiment!
Answer:
Determine peroxide number:
2KMnO4 + 5H2O2 + 3H2SO4  2MnSO4 + K2SO4 + 8H2O + 5O2
MnO4- + 5e + 8H+  Mn2+ + 4H2O
H2O2  2H+ + O2 +2e
2MnO4 + 5H2O2 + 6H+  2Mn2+ + 5O2 + 8H2O
Determine free fatty acids
CH3(CH2)14COOH (aq) + NaOH (aq)  CH3(CH2)14COONa (aq) + H2O
(l)

2. Mention that includes essential fatty acids for the body. Why is arachidonic
acid not an essential fatty acid?
Answer:
Linoleic acid (LA), an omega-6 fatty acid, and α-linolenic acid (ALA), an
omega-3 fatty acid, are considered essential fatty acids because they
cannot be synthesized by humans. Here is the classes of essential fatty
acid.

Page | 33
 Arachidonic acid is a polyunsaturated fatty acid with four cis double
bonds, which are the sources of its flexibility and give it the capacity to
react with molecular oxygen. Arachidonic acid is an omega-6 fatty acid. It
is one of the essential fatty acids that our body cannot manufacture.
Arachidonic acid is not one of the essential fatty acids. However it does
become essential if there is a deficiency in linoleic acid or if there is an
inability to convert linoleic acid to arachidonic acid which is required by
most mammals.
3. What is the difference between saturated and unsaturated fatty acids in the
oxidation process?
Answer:
If one or more of the bonds between the carbon atoms are double bonds,
the lipid is said to be “unsaturated”. If there is one double bond, the
triglyceride is said to be “monounsaturated”, if it has multiple double
bonds, it is “polyunsaturated”. Unsaturated fatty acids are usually liquid at
room temperature and are called oils. The double bonds in unsaturated
fatty acids can exzist in either a cis or a trans configuration. This describes
whether the hydrogen atom are on the same side (cis) or opposite sides
(trans). A cis double bond generates a bend in the molecule, influencing
its structure and downstream function.
4. What is the difference between oil and fat in terms of molecular structure?
Answer:

Page | 34
 Fats are molecules made up of a fatty acid chain with a glycerol head.
They're are several different kinds of fats determined by the number of
carbon atoms in the fatty acid chain and how many double bonds there are
in the chain and where the figure bonds are. While oils are made up of a
combination of different fats in varying concentrations. Both oil and fats
has general form triglyceride, it just differs in form. This difference is
based on differences in melting points because the molecular weight of oil
is greater than the molecular weight of fat. At room temperature, fat is solid
while oil is liquid.
L. References
Akhluwalia, V.K., Dhingra, S., and Gulati, A. 2005. College Practical
Chemistry. New Delhi: India Universities Press.
Carra, Sergio. 2018. Stepping Stoms to Synthetic Biology. Millan: Springer
Publisher.
Decker, Eric. A., Elias, Ryan J. and McClements, Julian D. 2010. Oxidation in
Food and Beverages. Cambridge: Elsevier.
Eaton, L and Rogers, Kara. 2017. Examining Basic Chemical Molecules.
London: Britannica.
Erickson, David R. 1990. Edible Fats and Oils Processing: Basic Principles
and Modern Practices. United States of America: Illinois.
Gordon, MH. 1990. The Mechanism of Antioxidants Action in Vitro. Dalam
B.J.F.Hudson, editor. Food Antioxidants. London: Elsevier.
Gunstone, F. D. 2012. Fatty Acid and Lipid Chemistry. United Kingdom:
Springer.
Gurr, M. I. 2012. Lipid Biochemistry: An Introduction. New York: Springer.
Insel, Paul M. 2014. Nutrition. Burlington: Jones and Barnett Learning.
Kent, Michael. 2000. Advanced Biology. Oxford: Oxford Univeristy Press.
Lawson, H.W. 2012. Standards for Fats and Oils. New York: Springer.
Lawson, H.W. 2013. Food Oils and Fats: Technology, Utilization and
Nutrition. New York: Springer.
Marcus, Jacqueline B. 2013. Cullinary Nutrition: The Science and Practice of
Healthy Cooking. Amsterdam: Elsevier.

Page | 35
McCance, Katthryn L. and Huether, Sue E. 2018. Pathophysiology: The
Biological Basis for Diseases in Adults and Children. New York:
Elsevier.
Moore, Barbara. 2013. Hydrogen Peroxide Health Benefits and Uses. New
York: Elsevier.
Rival, A. and Levang, P. 2014. Palm of Controversies: Oil Palm and
Development Challenge. London: Civor
Rodrigues, Thiago B. and Silva, Emiliano T. 2016. Molecular Diversity of
Environmental Prokaryotes. Boca Raton: CRC Press.
Rogers, Kara. 2011. The Human Body: Blood Physiology and Circulation First
Edition. New York: Britannica Educational Publishing.
Svehla, G. 1995. Vogel Buku Teks Analisis Anorganik Kualitatif Makro dan
Semimakro. Kalman Media Pustaka. Jakarta.

Page | 36
M. Attachment
1. Documentation
Tools and materials used

Tools used (erlenmeyer,

New cooking oil (yellow
beaker glass, graduated
solution) and used cooking oil H2SO4 4 N
cylinder, pipette, glass
(brownish yellow solution)
funnel)

PP indicator Aquades Alcohol 96%

Colorless buret
(colorless (colorless (colorless
and dark buret
solution) solution) solution)
a. Peroxide Number
1) Determination of the peroxide number of new oil
No Documentation Description
1 Measured 5 mL of new oil
(yellow solution) using
graduated cylinder 10 mL

Page | 37
No Documentation Description
2 5 mL of new oil was entered
into 2 erlenmeyer flask 100
mL

3 Measured 45 mL of aquades
using graduated cylinder 50
mL

4 45 mL of aquades entered
into the erlenmeyer

5 After added 45 mL aquades

the solution in the
erleenmeyer form two layer.
Top = yellow solution
Bottom = colorless solution

6 Measured 15 mL of H2SO4 4
N (colorless solution) using
graduated cylinder 50 mL

Page | 38
 No Documentation Description
7 45 mL of H2SO4 entered into
the erlenmeyer

8 The solution titrated with

KMnO4 solution ding dark
buret until the clor of the
solution change to be soft
pink

9 The solution after the

titration processs.
Top = yellowish solution
Bottom = soft pink solution

2) Determination of the peroxide number of used oil

No Documentation Description
1 Measured 5 mL of used oil
(brownish yellow solution)
using graduated cylinder 10
mL

Page | 39
No Documentation Description
2 5 mL of new oil was entered
into 2 erlenmeyer flask 100
mL

3 Measured 45 mL of aquades
using graduated cylinder 50
mL

4 45 mL of aquades entered
into the erlenmeyer.
After added 45 mL aquades
the solution in the
erleenmeyer form two layer.
Top = yellow solution
Bottom = colorless solution
5 Measured 15 mL of H2SO4 4
N (colorles solutiom) using
graduated cylinder 50 mL

6 45 mL of H2SO4 entered into

the erlenmeyer

Page | 40
 No Documentation Description
7 The solution titrated with
KMnO4 solution (dark puple
solution) using dark buret
until the color of the solution
change to be soft pink

8 The solution after the

titration processs.
Top = yellowish solution
Bottom = soft pink solution

b. Free Fatty Acid

1) Blank solution
No Documentation Description
1 Weighing 6 gram of aquades
using the analitical balance

Page | 41
No Documentation Description
2 6 gram of aquades entered
into the erlenmeyer flask

3 Measuring of 10 mL of
alcohol 96% using
graduated cylinder 10 mL

4 10 mL of alcohol 96% added

into the erlenmeyer, formed
colorless solution

5 3 dreps of PP indicator
added into the solution

6 Titrated with NaOH 0,1 N

(colorless solution) until a
pink solution is formed

Page | 42
 No Documentation Description
7 A pink solution formed after
titration.

2) Determination of the free fatty acid of new cooking oil

No Documentation Description
1 Weighing 6 gram of
new oil (yellow
solution), repeated
three times

2 6 gram of new oil

from each vial are
pout into three
different erlenmeyer

3 Measuring 10 mL of
alcohol 96 % using
graduated cylinder
10 mL

4 10 mL of alcohol
96 % added into each
erlenmeyer and
formed 2 layers
solution (top =

Page | 43
 No Documentation Description
yellow, bottom =
coloress)
5 3 dreps of PP
indicator added into
the solution

6 Titrated with NaOH

0,1 N (colorless
solution) until a pink
solution is formed

7 2 layers (top =
yellow and bottom =
soft pink) solution
formed after
titration.

8 Comparation of
titration resuli fot the
three erlenmeyer

3) Determination of the free fatty acid of used cooking oil

Page | 44
No Documentation Description
1 Weighing 6 gram
of used oil
(brownish yellow
solution), repeated
three times

2 6 gram of new oil

from each vial are
pout into three
different
erlenmeyer

3 Measuring 10 mL
of alcohol 96 %
using graduated
cylinder 10 mL

4 10 mL of alcohol
96 % added into
each erlenmeyer
and formed 2 layers
solution (top
=browinsh yellow,
bottom = coloress)

Page | 45
No Documentation Description
5 3 dreps of PP
indicator added into
the solution

6 Titrated with
NaOH 0,1 N
(colorless solution)
until a pink solution
is formed

7 2 layers (top =
brownish yellow
and bottom = soft
pink) solution
formed after
titration.
8 Comparation of
titration resuli fot
the three
erlenmeyer

Page | 46
2. Calculation
a. Determination of peroxide number
Known :
N KMnO4 = 0.1 N
V waste oil = 0.2 mL and 0.2 mL
N new oil = 0.1 mL and 0.1 mL
m
ρ=
v
m
0.89 g/mL=
5 mL
m=4.45 gram
Waste oil :
(0.2+0.2 )mL
v̅ = =0.2 mL
2
VKMnO4 x NKMnO4 x fp x 17
% H2O2 = x 100 %
5000
0.2 mL x0.091 x 1 x 17
= x 100 % = 0.0062 %
5000

In 5 mL of waste oil there are :

x
0.0062%= 5 x 100%
0.0062 x 5
x= = 3.1 𝑥 10−4 mL of H2O2
100

3.1 x 10-4 4.45

so, = 1000
x

x = 0.069 meq/kg
New oil :
(0.1+0.1 )mL
v̅ = =0.1 mL
2
𝑉𝐾𝑀𝑛𝑂4 𝑥 𝑁𝐾𝑀𝑛𝑂4 𝑥 𝑓𝑝 𝑥 17
% H2O2 = x 100 %
5000
0.1 𝑚𝐿 𝑥0.091 𝑥 1 𝑥 17
= x 100 % = 0.00309 %
5000

In 5 mL of waste oil there are :

𝑥
0. 00309% = 5 𝑥 100%
0.00309 𝑥 5
x= = 1.54 𝑥 10−4 mL of H2O2
100
1.54 𝑥 10−4 4.45
so, = 1000
𝑥

Page | 47
 x = 0.035 meq/kg
b. Determination Free Fatty Acid (FFA)
New cooking oil
V blank = 0.1 mL
V NaOH I = 0.2 mL
V NaOH II = 0.2 mL
V NaOH III = 0.3 mL
(𝑉𝑡𝑖𝑡𝑟𝑎𝑡𝑖𝑜𝑛 −𝑉𝑏𝑙𝑎𝑛𝑘 )𝑥 𝑁 𝑁𝑎𝑂𝐻 𝑥 𝐵𝑀 𝑝𝑎𝑙𝑚𝑎𝑡𝑖𝑐 𝑎𝑐𝑖𝑑
%FFA = 𝑥 100 %
𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 𝑥 1000

1st titration
(0.2 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.043%
2nd titration
(0.2 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.043%
3rd titration
(0.3 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.085%
(0.043 + 0.043 + 0.085)%
%FFA average = = 0.057 %
3

Waste cooking oil

V blank = 0.1 mL
V NaOH I = 0.3 mL
V NaOH II = 0.25 mL
V NaOH III = 0.3 mL
(𝑉𝑡𝑖𝑡𝑟𝑎𝑡𝑖𝑜𝑛 −𝑉𝑏𝑙𝑎𝑛𝑘 )𝑥 𝑁 𝑁𝑎𝑂𝐻 𝑥 𝐵𝑀 𝑝𝑎𝑙𝑚𝑎𝑡𝑖𝑐 𝑎𝑐𝑖𝑑
%FFA = 𝑥 100 %
𝑠𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝑔ℎ𝑡 𝑥 1000

1st titration
(0.3 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.085%

2nd titration

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 (0.25 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.064%
3rd titration
(0.3 −0.1) 𝑚𝐿 𝑥 0.1 𝑁 𝑥 2546.42 𝑔/𝑚𝑜𝑙
%FFA = 𝑥 100 %
6 𝑔𝑟𝑎𝑚 𝑥 1000

= 0.085%
(0.085 + 0.064 + 0.085)%
%FFA average = = 0.078 %
3

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