Q1-1

Experiment

English (Official)

Determination of Refractive Index of Highly Concentrated So-

lutions of NaCl and Glycerin using Total Internal Reflection
(TIR) (14 points)
Please read the general instructions in the separate envelope before you start this experiment.
Dependence of Refractive Index of a Solution on Concentration
Introduction:
Basic Principle: When a beam of light is incident on a surface, reflection and refraction take place. When
the surface is uneven (rough), these phenomena result in scattering light in all directions. If the surface
is that of a transparent medium, then most of the light is transmitted (refracted) and a small portion of
it is reflected. If the surface is opaque and polished (like a metallic surface), all the light is reflected.
 
 
In this experiment, the green laser light is shone normally into the water inside a container. The container
used has vertical walls. The laser beam first meets the smooth top water surface then it gets scattered
from the rough bottom surface of container while producing a bright green spot on the bottom surface.
This scattered light travels back into the water in all directions. This light again meets the top smooth
surface of water and undergoes reflection, refraction and another phenomenon called total internal
reflection (see Figure 1).
 
 
The scattered rays that reach the top surface of the water at an angle greater than the critical angle, get
totally internally reflected which results in a bright ring enclosing a dark region.
 

(a) Air (b) Liquid (c) Laser Pointer (d) Container with Liquid

Figure1: (Left) Arrangement to observe the phenomenon. (Right) The ray diagram.
 Q1-2
Experiment

English (Official)

From the definition of refractive index and critical angle we have:

1 √( 𝑑4 )2 + (ℎ)2 √(𝑑)2 + 16 × (ℎ)2

𝜇= = 𝑑 = − − − (1)
𝑆𝑖𝑛 (𝜃𝐶 ) 4
𝑑

Where µ is the refractive index (RI) of liquid, d is the diameter of the dark disc and h is the height/depth
of liquid. This formula can be applied to any transparent liquid medium.
 
From equation-(1)

(𝑉 )2
(𝑑)2 × (𝜇)2 = (𝑑)2 + 16 ×
(𝐴)2

Where, A is the effective area of the horizontal cross-section of the container and V is the volume. ℎ = 𝑉
𝐴

The diameter (d) as a function of refractive index and area of the container (A) is

4
𝑑= × 𝑉 = 𝑆 × 𝑉 − − − − − − − − − −(2)
𝐴 × √𝜇2 − 1

The diameter is proportional to volume and S is the proportionality constant given by

4
𝑆= − − − − − − − − − − − − − − − −(3)
𝐴 × √𝜇2 − 1

Using a liquid whose refractive index we know , we can calculate A. The effective area of the horizontal
cross-section of the container is given by

4
𝐴= − − − − − − − − − − − − − −(4)
𝑆 × √𝜇2 − 1

The present experiment is to determine the refractive indices of salt and Glycerin solutions at certain
high concentrations, using the refractive index of water (1.33).
 
Percent Concentration of Solutions:
Percent concentration volume per volume (V/V) is defined as the volume of solute in ml in 100 milliliters
of solution. Hence 50% solution of any solute is 50 ml of solute in 100 ml of solution).
 
 Q1-3
Experiment

English (Official)

Aim:
1. Determination of refractive index of 30% NaCl solution and Glycerin.
2. Determine the dependence of RI on the concentration of Glycerin water solution.

Equipments: You are supplied with the following equipment for this experiment:

Sr. No. Item Specifications Quantity

01 Green LASER Pointer Wavelength-532 nm 1 no + 1 spare
02 Burette stand As shown in final assembly 1 no
03 Beaker 500 ml 3 no
04 Syringe 50 ml 1 no
05 Digital thermometer To measure Room Temp 1 no
06 Glass stirrer For making solutions 1 no
07 Container As shown 1 no
08 Sodium Chloride (NaCl) Solution 30 % from AR grade salt 500 ml
09 Glycerin AR grade 500 ml
10 Distilled water Solution + washing 5000 ml
11 Tissue paper
12 Safety goggle Polaroid 1 no
13 Divider Screw adjustable 1 no
14 Steel scale (ruler – optional) 0.5 mm Least Count 1 no
15 Reading lens High quality 1 no

Warning :

Avoid direct eye exposure and through reflections.

Avoid staring at the laser spot for too long, advise to turn off the laser when it’s
not used for performing measurements

 
 Q1-4
Experiment

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The Experiment

Part-0 Measurement of the room temperature (0.2 points)

A.0 Measure the room temperature using the thermometer provided and record (0.2pt)
your reading in the answer sheet.
(Get the supervisor’s sign after taking this reading)

Steps for setting up the equipment .

Step-1: Making a cardboard box.

(a) Cardboard Holder (b) Cardboard Box (c) Tape

 Q1-5
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Wear your safety googles all the time. If you are already wearing spectacles, wear the safety
googles above that. Do not look directly into the laser light.
Switch off the laser light when you are not taking the readings.
Glycerin should be kept covered when not in use.

Part 1. Calculation of effective area of cross section (A) of the container using distilled
water (3.6 points)

Step-2: Attaching the Green LASER Pointer on the burette stand.

The laser pointer should be vertical .

(a) Cardboard Holder (b) Fisher Clamp (c) Laser (d) Opaque Rectangular container(TIR
Container) (e) Stand Rod
 Q1-6
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Step-3: Attaching the cardboard box on the burette stand.

(a) Cardboard Holder (f) Tape for support

 Q1-7
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Procedure for Observation:

Apparatus set up:
Clamp the laser light in the burette stand in such a way that the switch is pressed by the middle grip
of the stand. (When it is to be switched off, simply rotate the laser around the vertical axis so that the
pressure on the switch is released. When it is to be switched on, rotate it back to the original position).
Make sure that the base of the stand on which the container is placed is horizontal.
 
Switch off the laser light when you are not taking reading.
Place the container below the light so that the laser beam falls on the bottom of the container.
(i) Add 50 ml of distilled water into the container. Switch on the laser, a bright ring with a dark disc
inside becomes immediately visible.

A.1 (ii) Using the divider and ruler provided, measure the diameter of the dark (1.2pt)
disc. Use the reading lens for better observation of the disc diameter. Record
your readings in the Table 1 of the answer sheet.
(iii) Repeat steps (i) and (ii) by adding water in steps of 50 ml to obtain six
readings.
 

A.2 (iv) Plot a graph on the given graph sheet (Graph 1 - plot 1) with diameter of (1.8pt)
the dark disc (d) on the vertical axis and the volume (V) of water on the horizontal
axis using the given graph page in the answer sheet.
Note : (o,o) will be an additional data point to be plotted on the graph
(The total number of points to be plotted is 7)
while plotting use symbol
(.) dot = water
For marking the points on the graph
 

A.3 (v) Calculate the slope from the graph (S = d/V) (0.2pt)

A.4 (vi) Calculate the effective area (A) of cross-section of the container from the (0.4pt)
slope and equation (4)
 Q1-8
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Part 2: Determination of Refractive index of 30% NaCl Solution (3.4 points)

You are supplied 500ml of 30% NaCl Solution.
(i) Clean the container dry it by dabbing it with tissue paper.

B.1 (ii) Using the salt solution with fixed concentration, follow the steps (i) to (iii) of (1.2pt)
Part 1. Enter your readings in Table 2 of your answer sheet.
 

B.2 (iii) Plot a graph on the same graph sheet in the answer sheet (Graph 1 - plot (1.6pt)
2) overlapping graph plotted in Part 1) with diameter on the vertical axis and
volume on the horizontal axis. Label the points for distinction.
Note : (o,o) will be an additional data point to be plotted on the graph
(The total number of points to be plotted is 7)
while plotting use symbol
(+) plus = NaCl solution
For marking the points on the graph

B.3 (iv) Calculate the slope from the graph (0.2pt)

B.4 (v) Calculate the refractive index of 30% NaCl solution from the slope and the (0.4pt)
value of A calculated in Part 1.
 Q1-9
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Part 3-A: Determination of refractive index of Glycerin (3.4 points)

You are supplied with 500 ml of Glycerin.
(i) Clean the container and dry it by dabbing it with tissue paper.

C-1.1 (ii) Using the pure glycerin provided follow the steps (i) to (iii) of Part 1. Enter (1.2pt)
your readings in Table 3a of your answer sheet.
 

C-1.2 (iii) Plot a graph on the same graph sheet in the answer sheet (Graph 1 - plot (1.6pt)
3) overlapping graph plotted in Part 1 and 2) with diameter on the vertical axis
and volume on the horizontal axis. Label the points for distinction.
Note : (o,o) will be an additional data point to be plotted on the graph
(The total number of points to be plotted is 7)
while plotting use symbol
(*) star = Glycerin
For marking the points on the graph
 

(iv) Do not disturb this solution at this point as it is required for part 3B of this experiment.

C-1.3 (v) Calculate the slope from the graph. (0.2pt)

C-1.4 (vi) Calculate the refractive index of glycerin from the slope calculated in (0.4pt)
this part of experiment and the value of A already calculated in Part 1 of
this experiment.
 Q1-10
Experiment

English (Official)

Part 3B: Relation between Refractive index and concentration of Glycerin solution. (3.4
points)
Glycerin is miscible with water in all proportions; however, it takes thorough stirring to obtain a homo-
geneous mixture. In this part you will be measuring the refractive index of different concentrations of
aqueous solutions of Glycerin.
 
(i) Using the syringe provided, remove 150 ml of Glycerin from the container, so the remaining amount
of glycerin is 150 ml in the container.

C-2.1 (ii) Measure d and enter values of volume and diameter in Table 3b in the answer (1.6pt)
sheet.
(iii) Now add 50 ml of water to the container, stir the mixture gently and thor-
oughly to make
a homogeneous solution.
(iv) Calculate the new concentration of the solution.
(v) Measure the diameter of the ring and record values of volume, diameter and
concentration in Table 3b in the answer sheet.
(vi) Repeat steps (iii) to (v) for two more dilutions.

Calculate the values of S and Refractive Indices of the solutions and enter the values in the Table 3b in
the answer sheet.

C-2.2 (vii) Plot the values refractive index on the vertical axis against the concen- (1.4pt)
tration on the horizontal axis (graph -2) in the answer sheet.
Note : (o,1.33) will be an additional data point to be plotted on the graph .
(The total number of points to be plotted is 5)
 

At this stage you have measured the refractive indices of 30 % NaCl solution and glycerin. You have also
determined the relation between concentration and refractive index for glycerin solutions.
Answer the following questions in the answer sheet by choosing correct option:

C-2.3 How does the refractive index change with the concentration of glycerin (0.2pt)
solutions?
a. Increases with concentration
b. Decreases with concentration
c. Does not change with concentration

C-2.4 How would you expect the refractive index of NaCl solution to change with (0.2pt)
concentration?
a. Expected to increase with concentration
b. Expected to decreases with concentration
c. Expected not to change with concentration
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A1-1
English (Official)

Determination of Refractive Index of Highly Concentrated So-

lutions of NaCl and Glycerin using Total Internal Reflection
(TIR) (14 points)
Instruction – column on the right side is for office use only - do not write in this space

Part-0 Measurement of the room temperature(Get the supervisor's signature after taking
the temperature) (0.1 points)

A.0 (0.2 pt)

 
The room temperature is

Part 1. Calculation of effective area of cross section (A) of the container using distilled
water (3.4 points)

A.1 (1.2 pt)

Table 1:

S No. ...1... ...2... ...3... ...4... ...5... ...6...

Volume of water, V (ml)

Diameter of ring, d (cm)

A.2 (1.8 pt)

Graph 1.
 
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Graph-1
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A.3 (0.2 pt)

 
The slope from the graph (S = d/V)=
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.

A.4 (0.4 pt)

 
Effective area of cross section of the container (A) =
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.
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English (Official)

Part 2: Determination of Refractive index of 30% NaCl Solution (3.4 points)

B.1 (1.2 pt)

Table 2

S No. ...1... ...2... ...3... ...4... ...5... ...6...

Volume of solution, V (ml)

Diameter of ring, d (cm)

B.2 (1.6 pt)

Graph 1 Plot 2 (overlapping with previous Part).

B.3 (0.2 pt)

Slope from the plot 2 =
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.
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B.4 (0.4 pt)

Refractive Index of 30% NaCl solution
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.
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Part 3-A: Determination of refractive index of Glycerin at different concentrations (3.4

points)

C-1.1 (1.2 pt)

Table 3a:

S No. ...1... ...2... ...3... ...4... ...5... ...6...

Volume of solution, V (ml)

Diameter of ring, d (cm)

C-1.2 (1.6 pt)

Plot the graph in Graph 1 plot 3 (overlapping with previous Parts).

C-1.3 (0.2 pt)

Slope from the plot 3 =
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.
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C-1.4 (0.4 pt)

Refractive Index of glycerin=
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.
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Part 3B: Relation between Refractive index and concentration of Glycerin solution. (3.4
points)

C-2.1 (1.6 pt)

Table 3b:

S No. ...1... ...2... ...3... ...4...

Volume, V (cc)

Diameter, d (cm)

Concentration %

S= d/V (cm−2 )

R.I.

C-2.2 (1.4 pt)

Graph 2.
 
 
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Graph-2
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Calculations for concentrations

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.

Calculations for RI
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
.

At this stage you have measured the refractive indices of 30 % NaCl solution and glycerin. You have also
determined the relation between concentration and refractive index for glycerin solutions.
From your measurements, answer the following:
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C-2.3 (0.2 pt)

Choose the correct answer from the options given and fill in the blank
 
Refractive index of Glycerin........................
a. Increases with concentration
b. Decreases with concentration
c. Does not change with concentration

From the observations made above, give your prediction for NaCl solutions.

C-2.4 (0.2 pt)

Choose the correct answer from the options given and fill in the blank
 
Refractive index of NaCl solution..........................
a. Expected to increase with concentration
b. Expected to decreases with concentration
c. Expected not to change with concentration
 Q2-1
Experiment

English (Official)

Determination of the Glucose Content of Date Syrup Sample

( 8 Marks)
Please read the general instructions before you start this problem.
Date palms (Phoenix dactlylifera)are found in abundance in the Middle East and in desert areas.The latin
name of the tree is believed to have been derived from Greek Phoenix daktulos, which means purple
or red finger. In the UAE, the late Sheikh Zayed Bin Sultan Al Nahyan was the founder of the country’s
agricultural renaissance.He encouraged the people of UAE to cultivate dates beyond the borders of the
oasis where they traditionally grew dates, which played a great role in the flourishing dates throughout
the nation. This great achievement turned the desert into an abundant paradise for its dwellers; UAE
today brings this ancient super fruit to a new global market, which made the UAE the leader of date’s
cultivation in the world. From dates, a variety of food products are made.

Dates are rich in sugar which consists of two isomeric carbohydrates-Glucose (Molecular Formula
𝐶6 𝐻12 𝑂6 ) and Fructose (Molecular Formula 𝐶6 𝐻12 𝑂6 ). Glucose is the most important source of energy
in all organisms.
Glucose contains an aldehyde group (−𝐶𝐻𝑂) and hence is an aldose. Fructose contains a keto group
(−𝐶 = 𝑂) and hence is a ketose. Glucose can be quantitatively oxidized to Gluconic acid (𝐶6 𝐻12 𝑂7 ) by
iodine in alkaline medium.This enables the estimation of Glucose in presence by Fructose.
Date syrup is treated with iodine solution in the presence of 𝑁 𝑎2 𝐶𝑂3 solution.

2𝑁 𝑎2 𝐶𝑂3 (𝑎𝑞) + 𝐼2 (𝑎𝑞) + 𝐻2 𝑂 (𝑙) → 𝑁 𝑎𝐼 (𝑎𝑞) + 𝑁 𝑎𝑂𝐼 (𝑎𝑞) + 2𝑁 𝑎𝐻𝐶𝑂3 (𝑎𝑞)

 
𝐺𝑙𝑢𝑐𝑜𝑠𝑒 + 𝑆𝑜𝑑𝑖𝑢𝑚ℎ𝑦𝑝𝑜𝐼𝑜𝑑𝑖𝑡𝑒 ⟶ 𝑆𝑜𝑑𝑖𝑢𝑚𝐺𝑙𝑢𝑐𝑜𝑛𝑎𝑡𝑒 + 𝑆𝑜𝑑𝑖𝑢𝑚𝐼𝑜𝑑𝑖𝑑𝑒
Glucose: Sodium hypoiodite = 1:1
The iodate (I) (hypoiodite) is converted back to iodine after adding acid.
𝑁 𝑎𝐼( 𝑎𝑞) + 𝑁 𝑎𝐼𝑂( 𝑎𝑞) + 2𝐻𝐶𝑙( 𝑎𝑞) ⟶ 𝐼2 (𝑎 𝑞) + 𝐻2 𝑂( 𝑎𝑞) + 2𝑁 𝑎𝐶𝑙( 𝑎𝑞)
On the completion of the oxidation, the excess iodine is back titrated against 𝑁 𝑎2 𝑆2 𝑂3 solution using
starch as an indicator.
 Q2-2
Experiment

English (Official)

𝐼2 (𝑎𝑞) + 2𝑁 𝑎2 𝑆2 𝑂3 (𝑎𝑞) ⟶ 𝑁 𝑎2 𝑆4 𝑂6 (𝑎𝑞) + 2𝑁 𝑎𝐼 (𝑎𝑞) ( In acidic medium)

You are supplied with the following:

Chemicals Labeled as Quantity Supplied

Date Syrup Date Syrup Sample Supplied in Plastic Container
Iodine solution in a closed bottle Iodine 100 mL
0.10 M 𝑁 𝑎2 𝑆2 𝑂3 in a beaker 0.10 M𝑁 𝑎2 𝑆2 𝑂3 150 mL
15% 𝑁 𝑎2 𝐶𝑂3 in a beaker 15% 𝑁 𝑎2 𝐶𝑂3 50 mL
2M 𝐻𝐶𝑙 in a beaker 2M 𝐻𝐶𝑙 100 mL
Starch Starch Indicator 15 mL in Falcon Tube
Distilled Water Distilled Water 1000 mL in a bottle

Apparatus Labeled as Quantity supplied

25 mL burette on stand B1 1
10 mL volumetric pipettes P1 , P2 2
Pipette fillers 2
250 mL Glass stoppered bottles 3
150 mL conical flasks 𝐶1 to 𝐶3 3
Funnel 1
100 mL Volumetric flask V1 1
10 mL measuring cylinder 1
Wash bottle with distilled water 1
Dropper 1
Beaker 150mL 1

• 10 mL measuring cylinder is to be used for 15% Sodium Carbonate, 2M HCl and starch indica-
tor. Make sure that you wash it before every use.
• Feel free to use beakers, conical flasks and funnels that are available in your laboratory that
fit with the experimental requirements, if necessary.

Procedure
i)Standardization of Iodine solution
 Q2-3
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English (Official)

1. Rinse burette B1 with 3 to 5 mL of 0.1 M 𝑁 𝑎2 𝑆2 𝑂3

2. Fill burette B1 with 0.1 M 𝑁 𝑎2 𝑆2 𝑂3 using a funnel.
Note down the initial burette reading in Observation Table 1 in the answer sheet provided to you.
3. Using pipette P1 take exactly 10 mL iodine solution in a 150 mL conical flask 𝐶1 .
4. Using measuring cylinder add approximately 20mL of distilled water to the Iodine solution taken.
5. Titrate the iodine solution against 0.1M 𝑁 𝑎2 𝑆2 𝑂3 from burette B1 till a yellowish or light brown colour
appears.
6. Add 2 mL of starch indicator using 10 mL measuring cylinder , the solution turns blue, and continue
to titrate. The endpoint is colour change from blue to colourless.
7. Repeat the titration twice using conical flasks 𝐶2 . and 𝐶3 .

2.1 Record your titration readings in Observation Table 1 and note down the (3.0pt)
burette reading.

2.2 Calculate the molarity of Iodine solution. (0.5pt)

ii) Estimation of Glucose in Date Syrup

1. Using a funnel fill burette B1 with 0.1M 𝑁 𝑎2 𝑆2 𝑂3 .
2. Note down the initial burette reading in Observation Table 2 in the answer sheet provided to
you.
3. Dissolve the date syrup supplied in the plastic container with warm distilled water. Transfer the solu-
tion quantitatively in the 100 mL volumetric flask V1 and dilute with distilled water up to the mark using
dropper. (You can use funnel to transfer the glucose solution.)
4. Insert the stopper and shake the solution to homogenise it. Transfer the homogenised solution to a
correctly labelled 150 mL beaker.
5. Using pipette P2 , take exactly 10 mL of the homogenised solution in a 250 mL glass stoppered bottle.
6. Using a 10 mL measuring cylinder, add approximately 10 mL of 15% 𝑁 𝑎2 𝐶𝑂3 solution and add exactly
10 mL Iodine solution using pipette P1 .
7. Stopper the bottle and keep aside in the dark for about 30 minutes. Repeat Steps 5 and 6 using
two other 250 mL glass stopper bottles and keep them aside in the dark for about 30 mins each.
8. After 30 minutes, add 10 mL of 2M 𝐻𝐶𝑙 using measuring cylinder, stopper the bottle and shake well
(Iodine will liberate).
9. Add 𝑁 𝑎2 𝑆2 𝑂3 drop wise from the burette B1 till a light yellowish / light brown colour appears and
then add 2 mL of starch solution indicator (dark blue colour will be obtained) using a 10 mL measuring
cylinder and continue to titrate.
10. The endpoint is colour change from dark blue to colourless.
11. Repeat the titration twice.
 Q2-4
Experiment

English (Official)

𝐼2 (𝑎𝑞) + 2𝑁 𝑎2 𝑆2 𝑂3 (𝑎𝑞) ⟶ 𝑁 𝑎2 𝑆4 𝑂6 (𝑎𝑞) + 2𝑁 𝑎𝐼 (𝑎𝑞)

2.3 Record your titration readings in Observation Table 2 and note down the (3.0pt)
burette reading.
 
 
Molar mass of Glucose=180 g/ mol

Ask for the mass of Date syrup given to you m(g)=................................ g.

Observations
1.Calculate the number moles of Iodine solution added to 10 mL date syrup
=...........................
 
2. Calculate the number of moles of Iodine which remain unused in the glucose oxidation reaction
=
 
.......................................moles
 
 
3. Calculate the number of moles of Iodine used up for oxidation of 10 mL date syrup
 
 
 
=.......................................moles
 
4. Calculate the total number of moles Iodine used up for the oxidation of the given date syrup
 
 
 
 
 
the number of moles of Iodine =.......................................moles
 Q2-5
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English (Official)

2.4 Calculations (1.5pt)

 
Determine a) the number of moles b) mass c) percentage of Glucose present in
the given date syrup sample.
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A2-1
English (Official)

Determination of the Glucose Content of Date Syrup Sample

( 8 Marks)
Consider 1 Decimal place when recording your burette reading.

2.1 (3.0 pt)

Standardisation of Iodine Solution (Titration-1)
Observation Table 1
• 5 % deviation = 2.5 mark
• 10 % dev. = 1.5 mark
• 10-15% dev = 0.5 mark
• Consider the burette reading to one decimal place.

Sr. Titration Titration Titration

I II III

1 Initial burette reading mL

2 Final burette reading mL

3 Difference in burette readings mL

Burette reading =.......................................mL

(0.5 mark)

2.2 (0.5 pt)

Molarity of Iodine solution =......................................M
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2.3 (3.0 pt)

Estimation of glucose in date syrup (Titration-2)
Observation Table 2

Sr. Titration Titration Titration

I II III

1. Initial titration reading mL

2 Final Titration reading mL

3 Difference in burette readings mL

Burette reading =................................mL

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A2-3 English (Official)

2.4 (1.5 pt)

Calculations
1. Determine the mass of Glucose in the given sample of Date syrup. (1
mark total)
-Calculate the number if moles of glucose in the given sample (0.5 marks)
 
 
 
 
 
 
 
 
 
- Calculate the mass of glucose in the given sample (0.5 marks)
 
 
 
 
 
 
 
 
2.Determine the percentage of Glucose in the given sample of Date syrup.
(0.5 marks)
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Results:
1. Mass of Glucose in the given sample of Date syrup =...........................g
 
 
2. Percentage of Glucose in the given sample of Date syrup = .....................%
 Q3-1
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pH Titration using Universal Indicator (6 marks)

Please read the general instructions before you start this problem.
An acid base indicator is a substance that changes colour depending on the pH of the solution to which it
is added. Such substances are used in solution or powder form for the determination of pH of a solution
or to detect changes in pH during acid-base titrations.
A Universal indicator is able to change colours in a wide range of pH, and is hence used to determine how
acidic or basic a solution is. Universal indicator that can be used to indicate the pH of the solution. It is a
mixture of indicators that shows different colours at different pH values, and can be used in solution form
or as paper strips. A colour chart supplied with the indicator solution or strips enables the determination
of pH value.

pH range Description Colour

<3 Strongly acidic red
3 -6 Weakly acidic orange, yellow
7 Neutral green
8 – 11 Weakly alkaline blue
> 11 Strongly alkaline violet

For the ease of the translations use the following color index to express the color as a letter, so it is easier
for the markers to identify.

R Red
O Orange
Y Yellow
G Green
B Blue
V Violet

In case if you ONLY have Universal indicator as strips , use the following procedure. For every suc-
cessive additions from the burette , stir the solution well, then touch the solution with a glass rod or
dropper to take a very little of the solution and put it on the indicator strip and record the color.
Repeat this until the end of the titration.
 
Warning: This procedure is written so that error in using pH strips is minimised. However, it will
have an important error compared to the use of an indicator solution.

In this experiment, the equivalence point of an acid-base titration will be determined using Universal
indicator to determine pH during the titration of weak acid with strong base.
You are supplied with the following:
 Q3-2
Experiment

English (Official)

Apparatus Labelled as Quantity supplied

1 10 mL pipettes P3 ,P4 ,P5 and P6 4
2 25 mL burette B2 1
3 100 mL volumetric flasks V2 ,V3 ,V4 3
4 250 mL beaker 1
5 150 mL conical flasks C4 , C5 2
6 Funnel 1
7 Indicator bottles 2
8 Dropper* 1

* Use Dropper from Q-2

Chemicals Labelled as Quantity supplied

0.1M Succinic acid 0.1M Succinic acid 100 mL in a beaker
0.1M 𝑁 𝑎𝑂𝐻 (approx) 0.1M 𝑁 𝑎𝑂𝐻(approx) 100 mL in a beaker
Weak acid solution Weak acid In 100 mL volumetric flask V4
Universal indicator solution Universal indicator with colour chart In indicator bottle
Phenolphthalein indicator Phenolphthalein In indicator bottle
Distilled water Distilled water 1000 mL in a bottle

• Feel free to use beakers, conical flasks and funnels that are available in your laboratory that
fit with the experimental requirements, if necessary.
Procedure
1. Preparation of 100 mL of 0.01 M Succinic acid solution.
Using pipette P3 take 10 mL of the supplied 0.1 M succinic acid solution in volumetric flask V2 and dilute
up to the mark using distilled water.
2. Standardisation of the diluted 𝑁 𝑎𝑂𝐻 solution.
1. Using pipette P4 take 10 mL of the supplied 0.1 M(approx) 𝑁 𝑎𝑂𝐻 solution in volumetric flask V3 and
dilute up to the mark using distilled water.
2. Rinse burette B2 with 3-5 mL of diluted 𝑁 𝑎𝑂𝐻.
3. Using a funnel fill the burette with diluted 𝑁 𝑎𝑂𝐻 .
Note down the initial reading in Observation Table 1 in the answer sheet provided to you.
4. Using pipette P5 take 10 mL of 0.01M Succinic acid solution in the conical flask C4 and add 2 drops
phenolphthalein indicator.
5. Titrate the solution against the diluted 𝑁 𝑎𝑂𝐻 till a faint pink colour persists.
6. Repeat the titration until you get three reasonable readings.
 Q3-3
Experiment

English (Official)

3.1 Record your titration readings in Observation Table 1. Note down the read- (1.5pt)
ing.

3.2 Molarity of 𝑁 𝑎𝑂𝐻 solution used in titration = ....................M (0.5pt)

3. Titration of weak acid with strong base

1. Using a funnel fill burette B2 with diluted 𝑁 𝑎𝑂𝐻
2. Dilute the weak acid solution supplied in volumetric flask V4 to 100 mL using distilled water.
3. Using pipette P6 take10 mL of the diluted weak acid solution in a conical flask C5 and add 4 drops
Universal indicator.
In this titration, diluted 𝑁 𝑎𝑂𝐻 is added in instalments to the weak acid solution. After the addition of
each instalment of diluted 𝑁 𝑎𝑂𝐻, record the colour of the solution, and pH from the chart supplied to
you.
Put down pH value only when exact colour match is seen. If the colour is intermediate between
colours in the chart, write down the pH range.
4. Add the burette solution in instalments of 0.5 mL each till the colour of the solution changes to purple.
Thereafter take four more readings adding instalments of 0.5 mL each.

3.3 Record your observations in Observation Table 2 in the answer sheet pro- (2.5pt)
vided to you.

3.4 Plot a graph of pH vs volume of diluted 𝑁 𝑎𝑂𝐻 and determine the 5 mL (0.5pt)
range of the equivalence point.

3.5 Find ΔpH for every successive change in volume (0.5mL) and then plot a (0.5pt)
graph of ΔpH/ΔV vs volume of diluted 𝑁 𝑎𝑂𝐻 only in the range identified in
3.4 above

3.6 Determine the equivalence point from the data above. (0.5pt)
Experiment

A3-1
English (Official)

pH Titration using Universal indicator ( 6 Marks)

Consider 1 Decimal place when recording your burette reading.

3.1 (1.5 pt)

Observation Table 1
• 5 % deviation = full marks
• 10 % dev. = 1 mark
• 10-15% dev = 0.5 mark

Sr. Titration Titration Titration

I II III

1 Initial burette reading, mL

2 Final burette reading, mL

3 Difference in burette readings, mL

If only one reading has been taken, the mark will be 0.5

3.2 (0.5 pt)

Molarity of diluted 𝑁 𝑎𝑂𝐻 used for the titration = ....................M
Experiment

A3-2
English (Official)

3.3 (2.5 pt)

Observation Table 2

Volume of diluted Colour pH ΔpH ΔV Δ pH /Δ V

𝑁 𝑎𝑂𝐻 added in mL of solution

Observation Table 2 continued on the next page.

Experiment

A3-3
English (Official)

3.3 (cont.)

Volume of Diluted Colour pH ΔpH ΔV Δ pH /ΔV

𝑁 𝑎𝑂𝐻 added in mL of solution

Experiment

A3-4
English (Official)

3.4 (0.5 pt)

Graph of pH vs. volume of 𝑁 𝑎𝑂𝐻 added
Experiment

A3-5
English (Official)

3.5 (0.5 pt)

Graph of ΔpH/ΔV vs volume of 𝑁 𝑎𝑂𝐻 added
 
Experiment

A3-6
English (Official)

3.6 (0.5 pt)

Equivalence point=.................................. mL
 Q4-1
Experiment

English (Official)

General Instructions :
1. Wherever asked to mark the cell with a cross (X), mark as follows.

2. Wherever asked to mark the cell with a dash (-), mark as follows.

Q.4. Investigating mixup of newborn babies in a hospital

This is an investigative experiment to identify parents of three newborn babies who got mixed in a hos-
pital. The identification will be done with the help of blood groups. Untill the 1980s blood groups were
used in forensics, but this has now been replaced by other techniques that are more reliable. ABO blood
typing is still important for blood transfusions.
The presence or absence of A and B antigens on the Red Blood Cells (RBCs) determines the ABO blood
group of an individual. A person with only antigen A will be of blood group A. A person with only antigen
B will be of blood group B. A person with both antigens A and B will have blood group AB, while one who
has neither of the antigens has a blood group of O type. These antigens in an individual are governed
by the alleles of the gene responsible for their synthesis. Antigen A is determined by the allele 𝐼 𝐴 , while
antigen B is determined by the allele 𝐼 𝐵 . No antigens are produced when an individual carries the i allele.
The alleles 𝐼 𝐴 and 𝐼 𝐵 are co-dominant. The allele i is recessive to both 𝐼 𝐴 and 𝐼 𝐵 . alleles.
The presence of a given antigen can be identified by the use of antibodies. For example, if an antibody
against antigen A is added to blood from a person with blood group A, RBCs will clump together or
agglutinate. In this experiment, you will not use blood samples but solutions that mimic the process
of agglutination (clumping) of blood in the presence of a given antibody. The process has been mim-
icked using chemicals and precipitation represents the process of agglutination. The use of chemicals to
demonstrate the concept of blood groups was developed by Magdalena Wajrak of Edith Cowan Univer-
sity in Perth, Australia (Harrison T, 2015, Science in school, 32: 33-36)
Materials provided:
Glassware and miscellaneous items
1. Mimicked blood samples (13 in all) in 1.5 ml plastic tubes (in stand) labelled as follows:
(i). Four samples W, X, Y and Z (to be used in Exercise 1).
(ii). Nine samples C, D, E, 1F, 1M, 2F, 2M, 3F, 3M (to be used in Exercise 2).
2. 3 X 15 ml plastic tubes labeled as Anti – A, Anti - B and NA which have mimicked antibodies against
antigen A, antigen B and no antibody, respectively. These have been placed in a 250 ml plastic
beaker.
3. Four Cavity slides with 3 wells in each.
4. 7 plastic droppers.
5. 250 ml beaker with distilled water
 Q4-2
Experiment

English (Official)

6. Permanent marker
7. Rectangular labels
8. A4 sheet of black paper for placing slides
9. Waste bin
10. Additional distilled water in a bottle
Note: Please raise your hand, if you require additional distilled water.
Tissue papers and waste beaker will be provided by the supervisor
Exercise 1: Identify the blood groups of the blood samples W, X, Y and Z
1. Arrange four cavity slides to make a grid similar to the one shown in Figure 1.1. The slides should
be placed on the black sheet provided to you.

Figure 1.1

2. With the marker pen and labels provided, mark the wells of the grid as above.NA stands for no anti-
body.
3.1 Mark three droppers one as Anti-A (to be used only for taking Antibody A), one as Anti-B (to be used
only for taking Antibody B) and one as NA (to be used for taking the solution marked as NA).
3.2 Use the remaining droppers for blood samples.
3.3 Before starting and in between taking two different samples, flush the dropper several times (15 to
20 times) with distilled water to ensure that they are clean.
3.4 Ensure that the droppers are clean so that samples do not become cross-contaminated. Please avoid
touching the added solutions in any of the wells of the cavity slide.
4. Using a clean dropper, place a drop of blood sample W in each of the wells in column 1.
5. Continue the same with the other three blood samples (X, Y and Z).
6. In the first row, add 1 drop of Anti A (antibodies against A-antigen) in each of the wells.
7. In the second row, add 1 drop of Anti B (antibodies against B-antigen) in each of the wells.
8. In the third row, add 1 drop of NA solution.
 Q4-3
Experiment

English (Official)

A.1.1 Observe the wells and record the presence of white precipitate (mimicking ag- (0.75pt)
glutination of blood) in the appropriate cells in Table 1.1 by marking a cross
(X). Mark a dash (-) in cells representing wells where no precipitation was ob-
served.

Table 1.1
W X Y Z
Anti-A

Anti-B

NA

A.1.2 Request your supervisor to take a photograph of the plate. The supervisor will (0.25pt)
submit the photograph on your behalf.

Based on your observation, identify the blood groups of the samples W, X, Y and Z in Table 1.2 by marking
a cross (X) in the appropriate cell.

A.1.3 (0.25pt)

Table 1.2
Blood Group
Sample A B AB O
W

Identify the row(s) (Anti A, Anti B and NA) in Figure 1.1 that act(s) as the control for the experiment with
a cross (X) in the correct cell(s). Mark dash (-) in the remaining one(s).
 Q4-4
Experiment

English (Official)

A.1.4 (0.25pt)

Anti-A Anti-B NA

Exercise 2: Identify the blood groups of the parents and the babies in an attempt to restore the
babies to their respective parents.
There are three newborn babies (C, D and E), whose tags indicating their parents have been mixed up.
In order to identify the correct parents of the three babies, blood samples were taken from the babies
and the possible parents (1 to 3). The experiment attempts to identify the parents of the three babies
based on their blood groups.
For the identification you are provided with mimic of 9 blood samples labeled as follows:
Note: In the table, F stands for Father (not female) and M stands for Mother (not male).

Sample No. Label Blood sample from

1 1F Parent 1 father
2 1M Parent 1 mother
3 2F Parent 2 father
4 2M Parent 2 mother
5 3F Parent 3 father
6 3M Parent 3 mother
7 C Baby C
8 D Baby D
9 E Baby E

2.1. For each of the blood samples, identify the blood group following the procedure described in the
first exercise.
Note:
• Before reusing the cavity slides, wash them carefully with distilled water and dry them well with
tissue paper before putting samples in them.
• Ensure that the droppers are clean before taking the blood samples.
In Table 1.3, mark a cross (X) in the appropriate cells for the presence of precipitate. Mark a dash (-)
in cells representing wells where no precipitation was observed.
 Q4-5
Experiment

English (Official)

A.2.1.1 Take photographs of the labeled slides as done in 1.2. Request your supervisor (4.5pt)
to take photographs of the labeled slide. The supervisor will submit the photo-
graph on your behalf.

Table 1.3
1F 1M 2F 2M 3F 3M C D E
Anti-A

Anti-B

NA

Based on the results of the experiment, identify the blood groups of each of the nine samples by marking
a cross (X) in the appropriate cell in Table 1.4.

A.2.1.2 (0.50pt)

Table 1.4
Blood group of babies
Baby Blood group A Blood group B Blood group AB Blood group O
C
D
E
Blood group of parents
1F
1M
2F
2M
3F
3M

Based on the different blood groups identified by you, indicate the possible parent pairs for the babies
C, D and E by marking a cross (X) in the appropriate cell(s) in Table 1.5. There could be more than one
possibility. Mark a dash (-) in the remaining cell(s).
 Q4-6
Experiment

English (Official)

A.2.2 (1.0pt)

Table 1.5
Parent pair 1 Parent pair 2 Parent pair 3
Baby C
Baby D
Baby E

Based on your interpretations of the blood groups which baby(ies) (C, D, E) can be matched to their parent
pairs (1, 2, 3) with certainty based on the evidence?
Write the number of the parent pair (1, 2 or 3) in the cell to the corresponding baby(ies).
Mark a dash (-) in cell(s) corresponding to a child with multiple possible parent pairs.

A.2.3 (0.25pt)

Child C Parent
Child D Parent
Child E Parent

Predict the genotype of the child and the parent pair that can be matched with certainty based on your
answer in 2.3.
Indicate one possible genotype of the child and the corresponding parent pair.
Mark a dash (-) in cell(s) corresponding to a child with multiple possible parent pair.

A.2.4 (0.25pt)

Genotype Genotype of the parents

of the child Father Mother
Child C Parent
Child D Parent
Child E Parent
Experiment

A4-1 English (Official)

Q.4. Investigating mix up of newborn babies in a hospital

A.1.1 (0.75 pt)

Table 1.1

W X Y Z

Anti-A

Anti-B

NA

A.1.2 (0.25 pt)

Request your supervisor to take the photograph of the plate. The supervisor
will submit the photograph on your behalf.
Experiment

A4-2
English (Official)

A.1.3 (0.25 pt)

Table 1.2

Blood Group

Sample A B AB O

A.1.4 (0.25 pt)

Anti-A Anti-B NA
Experiment

A4-3 English (Official)

A.2.1.1 (4.5 pt)

Table 2.3

1F 1M 2F 2M 3F 3M C D E

Anti-A

Anti-B

NA

A.2.1.1 (cont.)

Request your supervisor to take the photographs of the plate. The supervi-
sor will submit the photograph on your behalf.
Experiment

A4-4 English (Official)

A.2.1.2 (0.50 pt)

Table 1.4

Blood group of babies

Baby Blood group A Blood group B Blood group AB Blood group O

Blood group of parents

1F

1M

2F

2M

3F

3M

A.2.2 (1.0 pt)

Table 1.5

Parent 1 Parent 2 Parent 3

Baby C

Baby D

Baby E
Experiment

A4-5
English (Official)

A.2.3 (0.25 pt)

Child C Parent

Child D Parent

Child E Parent

A.2.4 (0.25 pt)

Genotype Genotype of the parents

of the child Father Mother

Child C Parent

Child D Parent

Child E Parent
 Q5-1
Experiment

English (Official)

Q.5. Analyzing human chromosomes

The karyotype of a species represents the chromosomes of a cell, usually displayed as a
systematic arrangement of chromosome pairs in descending order of size. In order to
make a human karyotype, metaphase chromosomes are prepared from blood cells
The blood cells are induced to divide in culture and then treated with colchicine to block cell division
at metaphase. The cells are then broken by hypotonic treatment and chromosomes are spread on
glass slides. The chromosomes are stained and observed under a microscope. The photograph of the
metaphase (as shown in Figure 5.1) is then used to make a karyotype. Karyotypes can be used to identify
chromosomal abnormalities and aberrations. In the manual method, individual chromosomes are cut
from the photograph and then lined up by size and the position of the centromere with their respec-
tive partners. In humans chromosomes can be of three types, which are determined by the position
of the centromere (point of attachment of the mitotic spindle): (i) acrocentric chromosomes have the
centromere located very close to the end resulting into one of the arms being very short ( sometimes
not even discernable), (ii) submetacentric chromosomes have arms of unequal lengths and (iii) meta-
centric chromosomes have equal or almost equal arms. The karyotype prepared from the metaphase
spread shown in Figure 5.1 is presented in Figure 5.2. A description of the chromosomes belonging to
different groups (Figure 5.3) is given in Table 5.1.

Figure 5.1. A metaphase spread of human chromosomes

 Q5-2
Experiment

English (Official)

Figure 5.2. Karyotype prepared from the metaphase spread presented in Figure 5.1.

Table 2.1: Characteristics of the chromosomes in the human karyotype

Group Chromosomal Pairs Description
A 1-3 Large almost metacentric chromosomes
B 4-5 Large submetacentric chromosomes
C 6-12 + X Medium-sized submetacentric chromosomes
D 13-15 Large acrocentric chromosomes
E 16-18 Small submetacentric chromosomes
F 19-20 Small metacentric chromosomes
G 21-22 + Y Small acrocentric chromosomes
XY X Medium-sized sub metacentric chromosome
Y Small acrocentric chromosome

In the following exercise you are required to prepare a karyotype from the photograph of the metaphase
spread given to you. This chromosomal preparation is from an individual with an anomaly in their sex
chromosomes.
 
 Q5-3
Experiment

English (Official)

Materials given for karyotype preparation:

1. Photograph of a metaphase spread for karyotype preparation
2. Plastic petri-dish
3. Fine scissors
4. Forceps
5. Transparent tape
6. Sheet marked ‘Karyotype’ to paste the chromosome cutouts
Procedure:
Use the photograph of the metaphase spread for the following exercise
 
Exercise 1: Count the number of chromosomes
 

A.5.1 Count the number of chromosomes and record in the answer book. (0.25pt)
 Q5-4
Experiment

English (Official)

Exercise 2: Make a karyotype

1. With the help of fine scissors cut out each chromosome and place it in the given petri- dish. Make
sure that you do not loose any of them.
2. Arrange the chromosomes (cut outs) on the sheet labeled karyotype according to the size of the
chromosomes and position of their centromere. Use Figure 5.3 and Table 5.1 as your reference.
After arranging the chromosomes stick them in place with transparent tape. Take a photograph
and attach to the answer sheet.
3. The sheet labeled karyotype is part of your answer book and is placed at the end of the answer
book.
In each group, arrange the chromosomes by their approximate length. There will be no penalty for
errors in identification of specific chromosomes within a group.
Figure 5.3 is a pictorial guide on how to prepare a karyotype.

Figure 5.3 A pictorial representation of preparing a karyotype. 1: Items to be used for making
a karyotype, 2-5: Cutting the individual chromosomes, 6-8: Picking the chromosomes with the
fine forceps and collecting them, 9: Arranging the chromosomes, 10-11: Sticking the chromo-
somes with the piece of tape.

A.5.2 Please ask your supervisor to either scan or take a photograph of the karyotype (3pt)
on the answer sheet and submit.
 Q5-5
Experiment

English (Official)

Exercise 3: Answer the following.

Can the following cells in human blood be used for preparation of metaphase chromosomes?

A.5.3.1 Mark a cross (X) in the appropriate column (Yes / No). (0.25pt)

S.No. Cells Yes No

1. Erythrocyte (Red Blood Cells)

2. Lymphocyte (White Blood Cells)

You are asked to prepare chromosomal spread from mitotic plant cells. Can you use the following plant
parts to successfully prepare the chromosomal spread?

A.5.3.2 Mark a cross (X) in the appropriate cells (Yes/No). (0.25pt)

S.No. Plant parts Yes No

1. Leaf blade

2. Anther

3. Root tip

 
The photograph represents chromosomes in a rodent cell undergoing division. All chromosomes in this
rodent are acrocentric.
 Q5-6
Experiment

English (Official)

Does the following photograph represent the following stages of division?

A.5.3.3 Mark a cross (X) in the appropriate column (Yes/No). (0.25pt)

Stage of division Yes No

Mitotic Metaphase

Mitotic Anaphase

Meiotic Metaphase I

Meiotic Anaphase I

Meiotic Metaphase II

Meiotic Anaphase II
Experiment

A5-1 English (Official)

Q.5. Analyzing human chromosomes

A.5.1 (0.25 pt)

 
Number of chromosomes =

A.5.2 (3.0 pt)

 
Please ask your supervisor, to either scan or take the photograph of the kary-
otype ad submit.

A.5.3.1 (0.25 pt)

S.No. Cells Yes No

1. Erythrocyte

2. Lymphocyte

A.5.3.2 (0.25 pt)

S.No. Plant parts Yes No

1. Leaf blade

2. Anther

3. Root tip
Experiment

A5-2
English (Official)

A.5.3.3 (0.25 pt)

Stage of division Yes No

Mitotic Metaphase

Mitotic Anaphase

Meiotic Metaphase I

Meiotic Anaphase I

Meiotic Metaphase II

Meiotic Anaphase II
Experiment

A5-3
English (Official)

Karyotype