Govt College Women

University Sialkot
Department of Chemistry

LAB REPORTS
COURSE TITLE:
Analytical Chemistry (Lab-II)
SUBMITTED BY:
Group#04
Sidra Basharat Roll#13 Rida Tahreem Roll#91
Wajeeha Qaiser Roll#18 Hafiza Samra Roll#94
Maryam Basharat Roll#47 Zikra Shafiq Roll#95
Meezab Sarwar Roll#82 Soha Zulfiqar Roll#97
Hina Fatima Roll#90

SUBMITTED TO:
Dr. Iram Saba
SEMESTER:
8th (Analytical)
Submission Date
May 19, 2023
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 AIM:
To determine the strength of citric acid in lemon juice, by acid-base titration.

THEORY:
Citric acid is a weak organic acid occurring in many fruits, especially citrus fruits also found in many
fluids and tissues. It is very soluble and used as an additive in many drinks. The role of citric acid in drinks
is to improve taste and flavor, be antioxidant and maintain stability. Citric acid contains 3 carboxylic acid
groups and has a molecular formula of H3C6H5O7; 2-hydroxy-1, 2, 3-
propane tricarboxylic acid.
In human physiological blood, pH, and urine, it is found mainly as the
trivalent anions. Citrate salts are used to deliver minerals in
biologically available forms. In lemons and limes; citric acid is the
most concentrated compared with other fruits. Industries
manufacturing juice products must know the amount of juice, both for regulatory purposes (FDA) and for
their own manufacturing specifications. They can quantitatively
determine the amount of juice by measuring the amount of citric acid.
The procedure commonly used to do this takes advantage of the known
reactivity of citric acid with sodium hydroxide and is known as a titration

Acid-base titration: An acid–base titration is a method of

quantitative analysis for determining the concentration of an acid or base
by exactly neutralizing it with a standard solution of base or acid having
a known concentration. A pH indicator is used to monitor the progress
of the acid-base reaction.

BENEFITS OF CITRIC ACID:

Citric acid is commonly used as a food additive for natural flavoring and
as a preservative. It is also used for medical purposes, as an antioxidant.
1. Flavoring and preserving food: Citric acid can be added to processed and packaged foods and
drinks such as ice cream, sorbets, sodas, and wine. It is added as a preservative, emulsifying agent,
and flavoring. Citric acid is also added to many canned and jarred foods to help prevent botulism.
2. Antioxidant: Antioxidants, which are derived from citric acid, can help keep food edible over a
longer period. For example, sprinkling lemon juice, which contains citric acid, over apples or bananas
can help prevent them from turning brown. Ascorbic acid, better known as Vitamin C, is also found
in citric acid and is often used to help protect and preserve soft drinks and meats.

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 3. Medical: Citric acid is used to help kill harmful bacteria and infections on the skin’s surface that can
be common in people with diabetes, the elderly, and people who smoke. Citric acid also can be
combined with sodium citrate and potassium citrate to lower acid levels in the urine to help prevent
gout attacks.

EXPERIMENT

MATERIALS REQUIRED:
• Pipette
• Burette
• Funnel
• Conical flask
• Beaker
• Burette Stand
• Measuring cylinder
• China dish
CHEMICALS REQUIRED:
✓ NaOH solution
✓ Lemon juice (citric acid)
Figure: Sample solution of lemon juice
✓ Distilled water
Chemical Equation: C6H8O7 + 3NaOH Na3C6H5O7 + 3H2O
Standard Solution: 0.1M NaOH solution
Indicator: Phenolphthalein
End Point: Colorless to pink color
PROCEDURE:
1. Rinse the burette with (NaOH) solution and clamp it vertically in the burette stand.
2. Using a funnel, fill the burette (NaOH) solution. After that remove the funnel.
3. Note the initial reading of the burette.
4. Take a lemon and squeeze it to get lemon juice.
5. Filter the lemon juice to remove the pulp.
6. Prepare the lemon solution by taking 10ml of filtered lemon
juice into a 100ml flask and adding 90ml of distilled water.
7. Pipette out 10ml from this stock solution and transfer it to
the clean conical flask.
8. Then add 2-3 drops of phenolphthalein indicator.
9. Now add NaOH solution from the burette to the conical
flask, and swirl the flask continuously, until the color of the
solution in the flask changes to pink. Figure: pink color of the solution with indicator
10. Take 3 concordant readings and calculate the strength of
citric acid in lemon juice.
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 Figure: the pink End point of the solution
CALCULATIONS:
The volume of NaOH used = 11.63ml
Molar mass of citric acid = 192.124 g mol-1
(M1V1) citric acid = (M2V2) NaOH
M1 × 10ml = 0.1 × 11.63ml
M1 = 0.1 × 11.63
10
M1 = 0.1M
Strength of citric acid = Molarity × Molar mass
Strength of citric acid = 0.1 × 192.124
Strength of citric acid = 19.21gL-1

RESULT:
The strength of citric acid in lemon juice determined by acid-base titration is 19.21gL-1.

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 PRINCIPLE:
This method determines the Ascorbic Acid concentration in a solution by a redox titration using iodine.
Ascorbic acid more properly known as vitamin C, is an essential antioxidant needed by the human
body. As the iodine is added during the titration, the ascorbic acid is oxidized to dehydroascorbic acid,
while the iodine is reduced to iodide ions.

THEORY:
What is Redox Titration?
Redox titration is based on an oxidation-reduction reaction between the titrant and the analyte. It is one
of the most common laboratory methods to identify the
concentration of unknown analytes. redox titration involves the
transfer of electrons between the given analyte and the titrant.
An example of redox titration is the treatment of an iodine
solution with a reducing agent. The endpoint of this titration is
detected with the help of a starch indicator. In this example, the
diatomic iodine is reduced to iodide ions (I–), and the iodine
solution loses its blue color.
This titration is commonly referred to as iodometric titration.
TITRANT: whose concentration is known.
ANALYTE: whose concentration is to be determined.

What is Ascorbic Acid?

A vitamin that is found particularly in citrus fruits and green vegetables. It
is essential in maintaining healthy connective tissue and is also thought to
act as an antioxidant. Severe deficiency causes scurvy.
Commonly known as Vitamin C, has been found to neutralize free radical
molecules, which in excess can damage cells. Vitamin C is also involved
in the body's immune system by stimulating the activity of white blood
cells. Figure: Ascorbic Acid

HEALTH BENEFITS:

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It has been linked to many impressive health benefits, such as boosting antioxidant levels, lowering
blood pressure, protecting against gout attacks, improving iron absorption, boosting immunity, and
reducing heart disease and dementia risk.

EXPERIMENT

MATERIALS REQUIRED:
• Burette
• Tripod stand
• Conical flask
• Beakers
• Conical funnel
• Stirrer
• Measuring cylinder
• Pipette
• Whatt’s man filter paper no.1
REQUIRED CHEMICALS:
✓ 0.005mol/L Iodine solution
✓ Distilled Water
✓ Fresh Orange (Fruiter) Juice

Indicator: Starch is used as an indicator

Endpoint: When the color of the solution turns bluish black endpoint is reached.

PROCEDURE:
1. Take 10 ml of freshly prepared fruit juice from the above beaker with the help of a pipette.
2. Transfer it into a titration flask and add some drops of 0.5% starch
as an indicator.
✓ LEAKAGE TEST: Before performing titration, a leakage test
should be performed. Fill the burette with distilled water if there’s
any leakage then it can cause errors while taking readings.
3. Fill up the burette with iodine solution.
4. Put the above titration flask under the burette, open the stopper,
and perform titration.
5. Shake the flask frequently in order to see the color changes.
6. When the color of the solution turns bluish black endpoint is
reached. Figure: Orange Juice Sample

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7. Note down the reading. Repeat the same procedure three times and take three concordant readings.

Figure: Redox Titration Figure: Bluish Black Endpoint

CALCULATIONS:

Solution preparation
֍ Iodine solution:
→ In order to prepare (0.005 mol/L) Iodine solution, Weigh 2 g of potassium iodide into a 100 mL beaker.
→ Weigh 1.3 g of iodine and add it to the same beaker.
→ Now add some distilled water and swirl for a few minutes until the iodine gets dissolved.
→ Transfer the iodine solution to a 1000mL volumetric flask, making sure to rinse all traces of solution
into the volumetric flask using distilled water.
→ Make the solution up to the mark with distilled water.
֍ Starch indicator solution (0.5%):
Weigh 0.25 g of soluble starch and add it to 50 mL of near-boiling water in a 100 mL conical flask. Stir
to dissolve and cool before using.

֍ Fresh Fruiter Orange juice:

→ Cut the fruit into two halves, squeeze it in a
beaker to get some fresh juice.
→ Strain the juice using cheesecloth to remove
seeds and pulp that may otherwise block the
pipette. The stock solution is prepared.
→ Now Pipette out 10ml of fresh fruiter orange
juice from the stock into a separate beaker, fill
up the beaker with distilled water in order to
dilute it. The sample is ready for further
processing.

Figure: Orange Juice solution and iodine solution

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֍ READINGS:

Volume of Iodine used

Readings
from burette
1. 21ml
2. 23ml
3. 22ml

21+22+23
Volume used from burette =
3

= 22ml
Molar mass of Ascorbic Acid = 176.12 g/mol
M1V1 = M2V2
M1 × 10ml = 0.005mol/L × 22ml
0.005×22𝑚𝑙
M1 =
10𝑚𝑙

M1 = 0.011M
Strength of Ascorbic Acid = Molarity × Molar Mass
= 0.011 × 176.12 g/mol
= 1.93 g/L

RESULT:
The strength of Ascorbic Acid determined by redox titration in fruiter Orange is 1.93 g/L.

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 ANALYSIS OF DIFFERENT SAMPLES OF MILK:
The milk of different species although slightly varies in composition and
properties, contains the same constituents in general. On average, milk is made
up of 87.4% water and 12.6% milk solids (3.7% fat, 8.9% milk solids-not-fat).
The milk solids-not-fat contain protein (3.4%), lactose (4.8%), and minerals
(0.7%). Two major proteins present in milk are casein constitutes about 80% of
the total proteins and lacto albumin constitutes about 18%. A third protein
present in very small amounts in milk is lacto globulin.

AIM OF ANALYSIS:
The aim of this analysis is to analyze the Haleeb milk samples for composition and adulteration to have
awareness about its nutrition.

MATERIALS REQUIRED:
➢ Haleeb milk
➢ Test tubes
➢ Test tube rack
➢ Burette
➢ Burette stand
➢ Pipette
➢ Test tube holder
➢ Bunsen burner
➢ Beaker
➢ Gooch crucible
TESTS FOR MILK ANALYSIS:
1. Determination of proteins:
The presence of milk protein is determined by several tests that are discussed below:

A) BIURET TEST:
It is a chemical test that is used to see if an analyte has a peptide bond or not. As a result biuret test is
used to figure out how much protein is present in the analyte. The biuret test utilizes a biuret reagent that
is made up of a 1% solution of Copper II sulfate (CuSO₄), sodium hydroxide, and sodium-potassium

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tartrate. It is the Cu₂⁺ in the Biuret reagent that forms a chelate complex with the peptide bonds found in
protein resulting in a color change from blue to violet.

Biuret test
1. Take a cleaned and dried test tube.
2. Add 5 g of Milk sample in test tube.
3. Add 2ml of sodium hydroxide and 5 to 6 drops of copper sulfate
solution to it.
4. Shake the test tube gently to mix the ingredients thoroughly and
allow the mixture to stand for 4 – 5 minutes.
Interpretation
If bluish- violet color appears, it indicates the presence of protein in
milk.

B) XANTHOPROTEIC TEST:
The xanthoproteic test is used to detect amino acids containing an aromatic nucleus (such as tyrosine,
tryptophan and phenylalanine) in a protein solution which gives yellow color on heating with concentrated
sulfuric acid. The aromatic benzene ring undergoes sulfonation to give yellow colored product.

Xanthoproteic test
1. Take a cleaned and dried test tube.
2. Add 5 g of Milk sample in test tube.
3. Add a few drops of concentrated sulfuric acid and shake the
test tube.
4. Heat the test tube gently on a Bunsen burner.

Interpretation
If there is a formation of yellow precipitate, then the presence of
protein is confirmed.

C) MILLON’S TEST:
Millon’s test is specific to phenol-containing structures. Millon’s reagent is used to detect the presence of
soluble proteins. A few drops of the reagent are added to the test solution, which is then heated gently. A
reddish-brown coloration or precipitate indicates the presence of tyrosine residue which occurs in nearly
all proteins.
The reagent is made by dissolving metallic mercury in nitric acid and diluting it with water, forming
mercuric nitrate (Hg[NO3]2). In the test, the phenol group in the side chain of tyrosine gets nitrated, and
that product then complexes with Hg(I) or Hg (II) ions to give a red-colored precipitate.

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 Millon’s test Interpretation
1. Take a cleaned and dried test tube. If there is a formation of white precipitate
2. Add 5 g of Milk sample to test tube. or if the sample changes to brick red on
3. Add 2-3 drops of Millon’s reagent and shake well. heating then the presence of protein is
4. Observe the change. confirmed.

D) NINHYDRIN TEST:
This test consists of a chemical reaction that determines whether a sample compound contains amines or
alpha-amino acids. The main reactant in this process is ninhydrin, which is a hydrocarbon with the formula
C9H6O4. The marker for a positive ninhydrin test is a deep blue coloration obtained in the solution
indicating the presence of amino acids.

Ninhydrin test
1. Take a cleaned and dried test tube.
2. Add 5 g of Milk sample in test tube.
3. Add 1-2ml of ninhydrin solution into it and shake the test tube.
4. Boil the mixture and observe the change

Interpretation
If there is the appearance of deep blue or purple color then the
presence of protein is confirmed.

2. Determination of total solids by crucible method:

SNF or solids not fat is that nutrient portion present in milk which is other than milk fat and water. It
consists of protein (primarily casein and lactalbumin), carbohydrates (primarily lactose), and minerals
(including calcium and phosphorus. When SNF is combined with milk fat, then it is called total solids.

PROCEDURE:
1) Weigh the crucible.
2) Weigh 5 grams of milk in crucible.
3) Put crucible in water bath until dryness.
4) After complete dryness, put the crucible in oven and weigh after cooling.
5) Determine the percentage of total solids by using the formula.
CALCULATIONS:
(𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑐𝑟𝑢𝑐𝑖𝑏𝑙𝑒+𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒)𝑎𝑓𝑡𝑒𝑟 𝑑𝑟𝑦𝑖𝑛𝑔−𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑐𝑟𝑢𝑐𝑖𝑏𝑙𝑒
%age weight of total solid= 𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
× 100

Weight of crucible with lid = 25g

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Weight of milk sample before drying = 100g
Weight of milk sample after drying = 12g
% of total solids in milk sample = (25+12) -25/100 х 100
= 12%

Figure: Milk + Crucible Figure: Boiling Milk Figure: Total Solids in milk

RESULT:
The percentage of total solids present in 100g sample of Haleeb milk is 12%.

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 PRINCIPLE:
The titratable acidity is expressed as % lactic acid and is determined by titration of a known amount
of reconstituted milk with 0.1 N NaOH using phenolphthalein as an indicator. The amount of lactic
acid can be calculated by recording the volume of base required and the volume of the milk sample.
THEORY:
Raw milk quality is important to dairy farmers and processors because it affects the end product use and
economic value. Currently, raw milk quality is determined by fat, protein, total solids content, bacterial
counts, and somatic cell count. Titratable acidity is not one of the pay factors listed on the milk check
but has a strong economic impact because it is one of the criteria used to determine whether or not raw
milk enters the food chain as a premium-priced fluid product.

Titratable acidity (TA) is a rapid test (90 seconds to perform) indicating raw milk quality and
indirectly measuring the acid content in milk. Generally, as milk acid content increases, TA values
increase. All milk has a base acid content attributed to proteins, minerals, and dissolved gasses. Milk
acid content is increased by the bacteria that convert lactose to lactic acid. When this occurs, a dramatic
increase in TA value is observed. At the same time, milk has a strong buffering capacity (resisting a
change in the acid or alkali content) because of its protein content. Because these proteins resist a
change in the acid or alkali content, they, too, contribute to the “acidity” of milk.

Titratable acidity has been used for many years to indicate whether milk has undergone bacterial
degradation (acid production) or temperature abuse or is aged. Because raw milk refrigeration is
mandated by law, bacterial degradation and temperature abuse are no longer as prevalent as they once
were. Thus, TA values are fairly predictable, and high-quality raw milk has a relatively steady TA value
ranging between 14 to 17% (expressed as lactic acid). Today, two major factors impact the TA of raw
milk: age and protein content. Raw milk can be several days old before it is processed because
manufacturing centers are larger and fewer in number. Thus, as milk ages, bacteria grow and
subsequently decrease raw milk quality. Dairy cows are selected for increased milk protein content that
tends to increase the TA value such that the TA range of acceptable raw milk may change. Thus, it is
appropriate to study factors that have the greatest effects on the TA of raw milk.

 The titratable acidity test is employed to ascertain if milk is of such a high acidity as to reduce its
keeping quality and heat stability. The acidity of milk is of two kinds.
i. Natural acidity which is due to citrates and phosphates present in the milk and dissolved CO2 during
the process of milking and thereafter.
ii. Developed acidity which is due to lactic acid produced by the action of bacteria on lactose in milk.

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 Generally, the acidity of milk means the total acidity (Natural + developed) or titratable acidity. It is
determined by titrating a known volume of milk with standard alkali (a strong base), sodium
hydroxide, to the point of an indicator like phenolphthalein and expressing the results as % lactic
acid or in another traditional degree of acidity unit, such as Therner degrees (Th), Sohxlet-Henkel
degrees (SH), or Dornic degrees (D).
 The titratable acidity test measures the amount of alkali required to change milk’s pH from its initial
value of about 6-6 to 6.8, to the pH of the color change of phenolphthalein added to milk to indicate
the endpoint (pH 8.3). In fact, the method measures the buffering capacity of milk and not the true
acidity.
 This method is applicable to milk, cream, dried milk, condensed milk, evaporated milk, and simple
vanilla ice cream, whether neutralized or not. It is not applicable to chocolate ice cream, and some of
the fruits and other flavoring materials might contain ether-soluble materials that might interfere.

EXPERIMENT

APPARATUS REQUIRED:
▪ 10 ml pipette
▪ Beaker
▪ Measuring cylinder (25 ml).
▪ Burette (0.1 ml graduations).
▪ Burette stand
▪ Glass rod for stirring (flattened at one end).
▪ Weighing Balance
▪ 100 ml Erlenmeyer flask

REAGENTS:
✓ Milk Sample (Haleeb Milk)
✓ Phenolphthalein indicator Figure: Haleeb Milk Sample
✓ 0.1N sodium hydroxide solution

PROCEDURE:
1. Fill the burette with 0.1N NaOH solution.
2. Mix the milk sample thoroughly by avoiding the
incorporation of air.
3. Transfer 10 ml of milk sample with the pipette in a
conical flask.
4. Add 20 ml of glass distilled water to the same flask.
5. Add 1 ml of phenolphthalein indicator solution and stir
with a glass rod.
6. Take the initial reading of the alkali in the burette at the
lowest point of the meniscus.
Figure: Milk Sample with indicator

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7. Rapidly titrate the milk sample with 0.1N NaOH solution continue to add alkali drop by drop and stir
the content with a glass rod until a faint pink color is observed which remains constant for 10 to 15
seconds.
8. Complete the titration within 20 seconds.

Figure: Faint pink color of the Milk Sample solution

9. 8 ml of 0.1N NaOH solution was used for titration. Note down the final burette reading.
10. Calculate the percent acidity of the sample as lactic acid.
CALCULATIONS:
Solution preparation

֍ 0.1N NaOH:
𝑵𝒐. 𝑶𝒇 𝒆𝒒𝒖𝒊𝒗𝒆𝒍𝒆𝒏𝒕𝒔
• Normality =
𝑽𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝒔𝒐𝒍𝒖𝒕𝒊𝒐𝒏 𝒊𝒏 𝑳𝒊𝒕𝒓𝒆
No. of equivalents = Normality× Volume in L
= 0.1N × 1L
No. of equivalents = 0.1
𝑴𝒂𝒔𝒔 𝒊𝒏 𝒈
• Equivalents =
𝒆𝒒𝒖𝒊𝒗𝒂𝒍𝒆𝒏𝒕 𝑴𝒂𝒔𝒔
𝑴𝒐𝒍𝒂𝒓 𝒎𝒂𝒔𝒔
• Equivalent mass =
𝑵𝒐. 𝒐𝒇 𝒓𝒆𝒑𝒍𝒂𝒄𝒂𝒃𝒍𝒆 𝒔𝒑𝒆𝒄𝒊𝒆𝒔

Molar mass of NaOH= 40g/mol

NaOH→ Na+ + OH-
40
Equivalent mass = = 40g/mol
1
Mass in g = No. of equivalents × Equivalent mass
Mass in g = 0.1× 40g/mol
Mass in g = 4g

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→ It means that 4g of NaOH are required to prepare 0.1N solution of NaOH in 1000ml.

֍ Determination of % Lactic Acid:

The more sodium hydroxide added, the more acidic the milk. You can calculate the titratable acidity (as
lactic acid) as follows:
%Lactic acid = Volume of NaOH used × 0.09
= 8ml × 0.09
= 0.72%
Another formula to calculate titratable acidity is as follows:

Where,
V1 = volume in ml of the standard sodium hydroxide required for titration;
N = normality of the standard sodium hydroxide solution, and
V2 = volume in ml of milk taken for the test
9×8𝑚𝑙×0.1𝑁
%Lactic acid =
10𝑚𝑙
7.2
=
10
= 0.72%
OBSERVATION AND DISCUSSIONS:

Normal milk acidity ranges from 0.10 to 0.20% lactic acid. Any value in excess of 0.20 % can safely be
reckoned as developed lactic acid. The great variation in the original acidity shows the unreliability of
the titratable acidity as an indication of the presence of small amounts of developed acidity in unknown
samples. Due to the opacity of milk, the end-point of titration is not sharp, so care has to be taken to
adjust the conditions to reach the same end-point.

RESULTS:
Our results indicate that Habeeb milk sample contains 0.72% lactic acid, the amount present is over
about 0.002 per cent. A qualitative test showing the presence of more than this amount would indicate
souring. A quantitative determination of lactic acid would serve as an indication of the degree of
souring.

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 AIM:
The aim of this experiment is to determine the iodine value in edible oil.
Iodine value: The iodine value (IV) indicates the degree of unsaturation of a fat or oil. It is defined as the
number of grams of iodine absorbed by 100 g of fat. It is constant for a particular oil or fat.
Principle of iodine value: Under carefully monitored circumstances, the vegetable oil sample reacts with
an excessive amount of iodine monochloride solution (Wijs reagent). In unsaturated fatty acids, primarily
oleic and linoleic acids in the case of corn oil halogens quantitatively contribute to the double bonds.
Titrating with thiosulfate is used to determine unreacted halogens. The amount of halogen given as iodine
in grams that would react with 100 grams of oil is known as the “iodine number.”
THEORY:
Iodine value, also called iodine number, in analytical chemistry, measure of the degree of unsaturation
of an oil, fat, or wax; the amount of iodine, in grams, that is taken up by 100 grams of the oil, fat, or wax.
Saturated oils, fats, and waxes take up no iodine (since they have no double bond in their structures), and
therefore their iodine value is zero, but unsaturated oils, fats, and waxes take up iodine. (Unsaturated
compounds contain molecules with double or triple bonds, which are very reactive toward iodine.)

The more iodine is attached, the higher is the iodine value, and the more reactive, less stable, softer, and
more susceptible to oxidation and rancidification is the oil, fat, or wax. Iodine value is a useful parameter
in studying oxidative rancidity of oils since higher the unsaturation the greater is possibility of the oils to
go rancid .In performing the test, a known excess of iodine, often in the form of iodine monochloride, is
allowed to react with a known weight of the oil, fat, or wax, and then the amount of iodine remaining
unreacted is determined by titration.

֍ Why coconut oil is used as cooking oil and flaxseed not?

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Flaxseed oil for instance, has iodine value of roughly 170-200, signifying that it contains a high amount
of unsaturated fatty acids, therefore little saturated fatty acids. This high iodine value is correlated with
low stability and can pose a problem as it is more susceptible to oxidation and free radical production,
thus shortening its shelf life. This is why flaxseed oil is never used in cooking, since heat promotes readily
oxidation and polymerization. Polymerization constitute an irreversible process, causing the fatty acids to
solidify and become hard. On the other hand, coconut oil has the lowest iodine value range of 5-10. This
low unsaturation value makes it extremely resistant to oxidation and polymerization and thus a very safe
oil to store long term and cook with.

֍ Significance of iodine value in food industry:

The significance of iodine value within the food industry is that it acts as control, and quality assurance
parameter in oil products (for both edible and nonedible products).

EXPERIMENT

APPARATUS:
• Beakers
• Burette
• Conical flask
• Volumetric flask
• Tripod stand
• Pipette
CHEMICALS REQUIRED:
✓ Haali oil
✓ 0.1N Sodium thiosulphate
✓ Wijs solution
✓ Carbon tetra chloride (CCl4)
✓ Potassium iodide (KI)
✓ Starch solution
PROCEDURE:
1. Take 2.7 g of haali oil in sample flask.
2. Pour 25ml of CCl4 in a sample flask and pour 25ml of CCl4 in a
blank flask.
3. Now add 25ml of Wijs solution in a sample flask and close the
flask with stopper and same add 25ml of Wijs solution in a
blank flask and close the flask with stopper.
4. Shake well both.
5. Now add KI on the sides of stopper in both flasks.
6. Keep the flask for 30 minutes.
Figure: Haali oil sample flask+ blank flask

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7. Now add 10mL distilled water from stopper (washing of stopper in
both sample & blank flask).
8. Then Titrate sample flask against 0.1N Sodium Thiosulphate in
burette. Note change in color. Then add 1ml of 1% starch solution in
sample flask when the color is becoming lighter during titration. Then
again resume the titration. Milky white color indicates the end point.
9. Now perform same titration procedure with blank solution and milky
white color will be end point.
֍ End point:
Blank solution turned milky at end of titration while inside sample
solution there formed 2 layers milky layer on top while light brown at the
bottom. Figure: Titration of oil sample flask

Figure: Milky endpoint

CALCULATIONS:
Solution preparation

֍ 0.1N Sodium thiosulphate:

2.5 g Sodium thiosulphate in 80ml distilled water, after 10 minutes add more water till 100ml.
֍ Wijs solution:
3.3g of iodine in 200ml glacial acetic acid and gently warm it.

READINGS:
Vb = volume of sodium thiosulphate used for blank = 15ml
Vs = volume of sodium thiosulphate used for sample = 35ml
Vb = volume of sodium thiosulphate used for blank
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Vs = volume of sodium thiosulphate used for sample
Ws = weight of oil
N = normality of sodium thiosulphate

𝟏𝟐.𝟔𝟗 × (𝑽𝒃 − 𝑽𝒔) × 𝑵

Iodine Value =
𝑾𝒔

12.69 × (15−35) × 0.1 𝑁

Iodine Value =
2.7𝑔

Iodine Value = 9.4

RESULT:
A specific iodine value of 9.4 for an edible oil indicates that 9.4 grams of iodine can be absorbed by 100
grams of the oil. It signifies the degree of unsaturation or the presence of double bonds in the fatty acid
chains of the oil.

SAFETY PRECAUTIONS:
 Carbon tetrachloride is a carcinogen in laboratory animals.
 Avoid breathing vapors or skin contact. Work in a well-ventilated hood but avoid dispersing to
atmosphere.
 Store the Wijs reagent in a glass-stoppered amber bottle. Prepare fresh reagent every 30 days.
 If the iodine number is to be characteristic of the oil, the sample should be substantially free of
moisture and foreign materials.
 Agitation must be vigorous to assure removal and titration of iodine dissolved in the carbon
tetrachloride.

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