The COSHH Regulations

The Control of Substances Hazardous to Health Regulations (1988)

The COSHH regulations impose a duty on employers to protect employees and others from
substances used to work which may be hazardous to health. The regulations require you to make
an assessment of all operations which are liable to expose any person to hazardous solids, liquids,
dusts, vapors, gases or micro-organisms. You are also required to introduce suitable procedures
for handling these substances and keep appropriate records. It is essential that the laboratory
supervisor or some other person in authority is responsible for implementing the COSHH
regulations.

Part of the above regulations are to ensure that the relevant Health and Safety Data Sheets
are available for all hazardous substances used in the laboratory. Any person using a hazardous
substance must be informed of the following:

Physical data about the substance

Any hazard from fire or explosion
Any hazard to health
Appropriate First Aid treatment.
Any hazard from reaction with other substances.
How to clean/dispose of spillage.
Appropriate protective measures.
Appropriate storage and handling.

Page | 4
 Water-Borne Infections

For example, the microscopic bacterium called Legionella pneumophila will feed on any
scale, rust, algae or sludge in water and will breed rapidly if the temperature of water is between
20˚C and 45°C. Any water containing this bacterium which is sprayed or splashed creating air
borne droplets can produce a form of pneumonia called Legionaries Disease which is potentially
fatal.

Legionella is not the only harmful micro-organism which can infect water but it serves as
a useful example of the need for cleanliness. Under the COSHH regulations, the following
precautions must be observed. Any water contained within the product must not be allowed to
stagnate, i.e. the water must be changed regularly in glass wares.

Any rust, sludge, scale or algae on which micro-organisms can feed must be removed
regularly, i.e. the equipment must be cleaned regularly.

Where practicable the water should be maintained at a temperature below 20°C or above
45°C. If this is not practicable then the water should be disinfected if it is safe and appropriate to
do so. Note that other hazards may exist in the handling of biocides used to disinfect the water.

A scheme should be prepared for preventing or controlling the risk incorporating all of the
actions listed above.

Further details on preventing infection are contained in the publication “The Control of
Legionellosis including Legionnaries Disease”- Health and Safety Series booklet HS (G) 70.

Page | 5
 Lab Note Book Rubric

Excellent (2) Satisfactory (1) Unsatisfactory (0) Score

1. All experiments are written
All experiments are
completely and results
written completely Experiments are
Completion interpreted.
and results incomplete.
2. Graphs, tables and
interpreted.
suggestions are mentioned.
Copy is submitted
Copy is submitted at least one at the end of the Submission do not
Timely
day before oral evaluation semester within comply with the
Response
(viva). given timeline given time domain.
from lab instructor.
All experiments are concluded At least all
with correct interpretation of experiments are Objective of
Comments / results and any discrepancy in concluded with performing
Suggestions procedure or equipment is correct experiment is not
pointed out and correction interpretation of clear.
proposed. results.
1. Sources of error are correctly
identified.
Errors are not
2. Proper tools e.g. regression At least procedural
Sources of identified and
analysis is used to quantify / equipment errors
Errors reasons are
the magnitudes of errors. are identified.
unknown.
3. Effect of errors on results is
correctly interpreted.
1. A standard format is
followed for writing all
experiments.
Haphazard
Organization 2. Index of performing all
presentation
/ experiments is available at Overall neat look is
lacking any co-
Presentation the start of the notebook maintained.
ordination between
of Contents along with the date of
contents.
performing the experiment.
3. Graphs, tables are captioned
properly and attached.
Total Scores

Page | 6
 Experiment Performance Rubric

Below average Unsatisfactory

Excellent (4) Good (3) Satisfactory (2) Score
(1) (0)
1. 90% at least. 1. 80% at least. 1. 75% at least. 1. Below 75%.
2. In case of 2. In case of 2. In case of 2. In case of
Attendance 100 % leaves, lab leaves, lab leaves, lab leaves, lab
work is work is work is work is not
compensated. compensated. compensated. compensated.
Always on time in Always on time in Always on time in Always on time On time in less
Punctuality 95% labs at least. 90% labs at least. 80% labs at least. in 75% labs at than 75% labs at
least. least.
1. Calibration
checks of
equipment.
1. Calibration
2. Rinsing and
checks of
washing of 1. Rinsing and
equipment.
apparatus. washing of No
Pre- 2. Rinsing and Clear
3. Clear apparatus. understanding of
Experiment washing of procedural
procedural 2. Clear any experimental
Activities apparatus. understanding.
understanding. procedural activities.
3. Clear
4. Preparation of understanding.
procedural
solutions /
understanding.
chemicals /
accessories for
experiment.
Active
participation in: Active
participation in:
1. Keeping
Active
workplace 1. Keeping
participation in:
clean. workplace Active
2. Collection of clean. participation in: No active
1. Keeping
Participation data. 2. Collection of participation in
workplace
3. Proper data. 1. Collection lab activities.
clean.
handling of 3. Proper of data.
2. Collection of
equipment / handling of
data.
chemicals. equipment /
4. Cooperating chemicals.
with team
members.
1. Proper Proper cleaning No participation
Post 1. Proper cleaning 1. Proper cleaning / washing of
cleaning / in post
Experiment / washing of / washing of items.
washing of experiment
Activities items. items.
items. activities.

Page | 7
 2. Safekeeping of 2. Safekeeping of 2. Safekeeping
items and items and of items and
accessories. accessories. accessories.
3. Proper waste 3. Proper waste
disposal of disposal of
chemical. chemical.
4. Proper shut
down of
equipment.
1. All possible
hazards are
evaluated.
2. Proper
1. All possible
functioning of 1. All possible
hazards are
safety hazards are
evaluated.
parameters is evaluated. All possible
2. Proper
evaluated. 2. Proper hazards are No possible
functioning of
Safety 3. PPE’s are used functioning of evaluated hazard evaluation
safety
properly. safety is done.
interlocks is
4. MSDS are interlocks is
evaluated.
consulted and evaluated.
3. PPE’s are
Lab safety form
used.
is filled
countersigned
by lab
instructor.
Result 1. Proper data is
Calculation collected from 1. Proper data is
experiment. collected from
2. Erroneous experiment. 1. Erroneous
readings / 2. Erroneous readings /
trends in data readings / trends in data All results are No results
are identified. trends in data are identified. calculated. calculated.
3. All results are are identified. 2. All results are
calculated. 3. All results are calculated.
4. Unit calculated.
consistency is
maintained.
Discussion Results are: Results are:
Results are:
on Results 1. Accurate 1. Accurate No idea has been
1. Accurate Results are
2. Precise 2. Precise built from data
2. Precise accurate.
3. Reproducible 3. Reproducible and results.
4. Presentable
Data / Results are Data / Results are Data / Results are
presented in: presented in: presented in:
Graph / Data / Results No graphical /
1. Graphical form. 1. Graphical 1. Graphical
Statistical are presented in statistical
2. Statistically form. form
Analysis of graphical form. manipulation of
analyzed e.g. 2. Statistically Statistically
Data data.
trend lines analyzed e.g. analyzed e.g.
regression trend lines trend lines

Page | 8
 coefficients regression regression
added. coefficients coefficients
3. Errors marked added. added.
graphically. 3. Errors marked
4. Graphs graphically.
properly
captioned.
1. Objectives of
experiment are
fully grasped.
1. Objectives of
2. Idea from
experiment are
experiment can
fully grasped. 1. Objectives of
be applied to
2. Idea from experiment are
other physical
experiment can fully grasped.
situations. Objectives of
Analytical be applied to 2. Idea from No idea is built
3. Any betterment experiment are
Thinking other physical experiment can from experiment.
to current fully grasped.
situations. be applied to
procedure is
3. Any betterment other physical
proposed.
to current situations.
4. One has
procedure is
developed
proposed.
theoretical
understanding
of concept.
Total Scores

Page | 9
 Equipment Status in Chemical Process Industries Lab

Max.
Place of
Sr. No. Equipment Remarks Recommendations Expected
Availability
Price

1 Oven Working Condition - - -

2 Muffle Furnace Working Condition - - -

Gives Erroneous Brandth

3 Weight Balance Purchase New Rs. 4000
Results Road.

Page | 10
 List of Chemicals for Chemical Process Industries Lab

Required Available
Sr. No. Chemicals Experiment
Quantity Quantity
EDTA Solution (Di-Sodium Salt of
1 Ethylene Diamine Tetra Acetic Acid Ex. 2 0 kg 3.7 kg
Na2H2C10H12O8N2.2H2O)
2 Murexide Indicator Ex. 2 0 kg 0.05 kg
3 Buffer Solution (NH4Cl + NH4OH) Ex. 2 0 kg 1 kg
4 Glucose Ex. 4 0 kg 1 kg
5 Copper Sulphate (CuSO4.5H2O) Ex. 4 0 kg 1 kg
Ex. 4, Ex. 5, Fresh Fresh
Ex. 7, Ex. 8, available available
6 Distilled Water
Ex. 9, Ex. on on
10, Ex. 11 demand. demand.
7 Rochelle Salt (Sodium Potassium Tartrate) Ex. 4 0 kg 1 kg
Ex. 4, Ex. 8,
8 Sodium Hydroxide (NaOH) 0 kg 1 kg
Ex. 11
9 Methylene Blue Indicator Ex. 4 0 kg 0.025 kg
Fresh Fresh
available available
10 Milk Ex. 4, Ex. 5
on on
demand. demand.
11 Acetic Acid Ex. 4, Ex. 5 0 kg Sufficient
Ex. 6 Fresh Fresh
available available
12 Leather
on on
demand. demand.
13 Potassium Hydroxide (KOH) Ex. 7, Ex. 12 0 kg Sufficient

Page | 11
 Ex. 7, Ex. 8,
14 Hydrochloric Acid (HCl) 2 kg 0 kg
Ex. 9, Ex 11
15 Phenolphthalein Indicator Ex. 7, Ex .8 0 kg 0.125 kg
Fresh Fresh
available available
16 Vegetable Oil Ex. 7
on on
demand. demand.
17 Ethanol Ex. 7, Ex. 8 2 liter 0 liter
Ex. 8 Fresh Fresh
available available
18 Soap
on on
demand. demand.
19 Stearic Acid Ex. 8 0 kg 0.5 kg
20 Methyl Orange Indicator Ex. 8, Ex. 11 0 kg 0.1 kg
21 Sodium Silicate (Na2SiO3) Ex. 9 0 kg 1 kg
22 Potassium Chromate (K2CrO4) Ex. 10 0 kg 1 kg
23 Silver Nitrate (AgNO3) Ex. 10 0g 500 g
24 Ammonium Per Sulphate (NH4)2S2O8 Ex. 10 0 kg 1 kg
25 Potassium Permanganate (KMnO4) Ex. 10 0 kg 1 kg
26 O-Phenanthroline Indicator Ex. 10 0g 10 g
27 Mohr’s Solution Ex. 10 0 kg 1 kg
28 Ferric Chloride FeCl3.6H2O Ex. 11 0 kg 1 kg
29 Potassium Ferrocyanide K4Fe(CN)6.3H2O Ex. 11 0 kg 0.5 kg
30 Glycerin Ex. 12 0 liter 1 liter
31 Red Oil (Oleic Acid) Ex. 12 1 liter 0 liter
32 Silica Gel Ex. 9 1 kg 0 kg
33 EBT (Eriochrome Black-T) Indicator Ex. 3 0 kg 0.1 kg
34 Sodium Carbonate (Na2CO3) Ex. 11 0 kg 0.5 kg

Page | 12
List of Glass / Metal Wares for Chemical Process Industries Lab

Required Available
Sr. No. Item Name Experiment
Quantity Quantity
1 Petri Dish Ex. 1 0 11
2 China Dish - 0 8
Ex. 1, Ex. 5, Ex. 6, Ex. 1
3 Weight Balance 1
7, Ex. 8 (Defected)
4 Conical Flask (500ml) - 0 3
Ex. 2, Ex. 3, Ex. 4, Ex.
5 Conical Flask (250ml) 0 13
7, Ex. 8, Ex. 10
Conical Flask with Nozzle
6 - 0 1
(250ml)
7 Conical Flask (125ml) - 0 1
8 Burette (Extra Large Size) - 0 1
Ex. 2, Ex. 3, Ex. 4, Ex.
9 Burette (50ml) 0 12
7, Ex. 8, Ex. 10
Ex. 2, Ex. 3, Ex. 4, Ex.
10 Pipette (25ml) 0 4
5, Ex. 7, Ex. 8, Ex. 10
11 Pipette (10ml) - 0 6
12 Small Hand Pipette - 0 24
13 Pipette Rubber Bulb - 0 1
Ex. 4, Ex. 5, Ex. 7, Ex.
14 Wire Gauze 10 2
10, Ex. 12
Ex. 4, Ex. 5, Ex. 7, Ex.
15 Tripod Stand 0 3
9, Ex. 10, Ex. 12
Ex. 4, Ex. 5, Ex. 6, Ex.
16 Burner 7, Ex. 9, Ex. 10, Ex. 0 3
12
17 Beaker (500ml) - 0 13

Page | 13
18 Beaker (400ml) - 0 8
Ex. 4, Ex. 5, Ex. 7, Ex.
19 Beaker (250ml) 8, Ex. 10, Ex. 11, Ex. 0 14
12
20 Beaker (100ml) - 0 6
21 Beaker (50ml) - 0 4
22 Beaker (20ml) - 0 2
23 Funnel (Extra Small) - 0 1
24 Funnel (Small) Ex. 4, Ex. 5, Ex. 11 24 6
25 Funnel (Medium Size) - 0 21
26 Whatman Filter Paper No. 41-42 Ex. 4, Ex. 5, Ex. 11 3 Packets 0
27 pH Paper - 2 Packets 0
28 Crucible with Lid Ex. 6
29 Holder Ex. 6 10 1
30 Desiccator Ex. 6, Ex. 8 0 3
31 Spherical Bottom Flask (6000ml) - 0 1
32 Spherical Bottom Flask (1000ml) - 0 1
33 Spherical Bottom Flask (500ml) Ex. 7 4 0
Spherical Bottom Three Nozzle
34 - 0 5
Flask (1000ml)
35 Flask Mouths - 0 9
36 Condenser of Flask Ex. 7 0 16
37 Glass Rod Ex. 8, Ex. 9 0 13
38 Test Tube Ex. 10, Ex. 12 0 40
39 Glass Dish - 0 13
Micro Filtration Unit Cylinder
40 - 0 1
(300ml)
41 Pyrex Bottle (2000ml) - 0 1
42 Pyrex Bottle (1000ml) - 0 22
43 Pyrex Bottle (500ml) - 0 12

Page | 14
44 Pyrex Bottle (250ml) - 0 6
45 Pyrex Bottle (100ml) - 0 4
46 Pyrex Bottle (50ml) - 0 1
47 Pyrex Bottle (25 ml) - 0 1
48 Separating Funnel (1000ml) - 0 1
49 Separating Funnel (500ml) - 0 2
50 Separating Funnel (125ml) - 0 4
51 Separating Funnel (50ml) - 0 1
52 Glass Jar (500ml) - 0 1
53 Glass Cylinder (250ml) - 0 1
54 Glass Cylinder (100ml) - 0 3
55 Glass Cylinder (25ml) - 0 4
56 Glass Cylinder (10ml) - 0 8
57 Crucible (50ml) - 0 18 + Extra
58 Crucible (25ml) - 0 18 + Extra
Chemical Storage Bottle
59 - 0 1
(1000ml)

Page | 15
 Lab Manuals (Experiments)

1. General Safety Precautions

ANALYSIS BASED EXPERIMENTS

2. Experiment 1
(Analysis of Total Solids of Water)
3. Experiment 2
(Analysis of Calcium Hardness of Water)
4. Experiment 3
(Analysis of Total Hardness of Water)
5. Experiment 4
(Analysis of Sugar in Milk)
6. Experiment 5
(Analysis of Fat in Milk)
7. Experiment 6
(Analysis of Leather)
8. Experiment 7
(Analysis of Vegetable Oil)
9. Experiment 8
(Analysis of Soap)
10.Experiment 9
(Analysis of Silicate)
11.Experiment 10
(Analysis of Chromate)
12.Experiment 11
(Analysis of NaOH)
13.Experiment 12
(Analysis of Biomass)

SYNTHESIS BASED EXPERIMENTS

12.Experiment 13
(Synthesis of Prussian Blue)
13.Experiment 14
(Synthesis of Liquid Soap)

Page | 16
 General Safety Precautions

Precautions: (Common for all experiments)

1. Personal Safety:
 Use fire extinguishers in case of fire or explosion.
 Do not smoke because flammable liquids, gases, and vapors can cause fire.
2. Equipment Safety:
 Do not exceed upper limits of operating conditions (T = Temp., P = Pressure, F
= Flowrate, S = Speed).
 Check proper working of equipment ON/OFF Circuit Breaker or Switch.
 Do not use equipment without lab attendant.
 Do not try to become a juggler with equipment but if you don’t know anything
then report and ask for it.
3. Chemical Safety:
 Drain any residual water present in the apparatus.
 Do not expose flammable organic fluids with flame or spark.
 Do not eat any food in chemical laboratory because its contamination with any
dangerous chemical can cause death.
 Do not taste any chemical it can cause death.
 For dilution purposes of acids and alkalis, the acid or alkali should be added
slowly drop by drop into water while stirring. The reverse process surely causes
explosive phase change of water. (e.g. H2SO4 dilution).
 Gloves and Goggles must be worn whenever there is a risk to the eyes.
 Wear lab coat.

Page | 17
ANALYSIS BASED EXPERIMENTS

Page | 18
 Experiment 1
(ANALYSIS OF TOTAL SOLIDS OF WATER)

Objective:

Estimation of total solids in given sample of tap water.

Equipment Setup / Apparatus:

Oven, Petri Dish, Weight Balance

Oven and Weight Balance

Page | 19
Reagents:

Tap Water

Theory:

For any ordinary analysis of water the following determinations are made:

1) Chemical Tests: Total Solids, Alkalinity or Acidity, PH, Chlorides, Sulphates,

Nitrates, Phosphates, Iron, Aluminum, Calcium and Magnesium Hardness, Free Saline
and Ammonia, Silica, Dissolved Oxygen, Oxygen Consumed, Free Carbon dioxide,
Poisonous Metals like Pb, As, Zn, Ba, Sr, Mn etc.
2) Physical Tests: Odor, Color, Taste, Turbidity, and Suspended Matter.

The total solid contents consist of:

a) Insoluble Solids
b) Soluble Solids
c) Volatile Matter
d) Residue

Almost same method is used for the determination of all types of above solid contents.
Here Gravimetric Method is employed for the measurement of solid contents.

Procedure:

1) A petri dish was cleaned, dried, and weighed properly.

2) Volume of 25ml of Tap Water were poured in the dish.
3) This dish along with water was placed in the oven at 103oC.
4) Duration of heating will be an hour or longer period to attain constant weight.
5) The water was evaporated in this way.
6) The dish was weighed again after desiccation.
7) Then the results were calculated.

Data Analysis:

Initial Weight (Weight of Empty Petri Dish) = X _______________ grams

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Final Weight (Weight of Petri Dish + Solids) = Y ________________ grams
Increase in Weight (Weight of Total Solids) = Y-X = Z ______________ grams
Volume of Sample = 25 ml
× 1000000
( )=

Page | 21
 Experiment 2
(ANALYSIS OF CALCIUM HARDNESS OF WATER)

Objective:

Estimation of calcium hardness in water sample.

Equipment Setup / Apparatus:

Conical flask, Burette, Pipette, Iron stand, Oven.

Page | 22
 Oven

Reagents:

Hard Water, EDTA Solution, Murexide Indicator, Buffer Solution

Theory:

Water becomes hard because of the presence of soluble salts such as Bicarbonates,
Sulphates, Chlorides of Calcium, and Magnesium and also because of carbonates of magnesium.
Hardness may be called as carbonate hardness or non -carbonate hardness. The carbonate and
bicarbonate hardness is known as temporary hardness, whereas that of chlorides and Sulphates is
known as permanent hardness. Hardness causes problems in washing/cleaning and steam raising
by boilers. For drinking purposes water, should be soft but hard water can also be used. Temporary
hardness can be removed just by boiling but permanent hardness cannot be removed by boiling.

Hardness measurements can be made by gravimetric as well as by titrimetric methods. In

the gravimetric method, calcium as precipitates of calcium oxalate/carbonate is obtained. While
Page | 23
magnesium is weighed as magnesium pyrophosphate. Titrimetric method being more accurate and
time saving, so it is to be used for the determination of the hardness. In these titrations buffer
solution is to be used to keep the pH constant during titration.

1. BUFFER SOLUTION
A buffer solution is a weak acid and a salt of that acid with strong base. The most favorable
buffer solution for this titration is prepared by using the mixture of NH4Cl/NH4OH having
PH=10. For this buffer solution take 70g of NH4Cl and add 568ml of conc. NH3OH
Solution and then stir and dilute the solution to 1 Liter with deionized water.
2. EDTA / Standard Solution / 0.01 M
EDTA is the Di-Sodium Salt of Ethylene Diamine Tetra Acetic Acid. Dry 5 g of Di-Sodium
Di-Hydrogen ethylene diamine tetra acetic acid (Na2H2C10H12O8N2.2H2O) in oven at 80oC
for 1.5 hr. From dried salt take 3.7224g in water, dissolve it and make up to 1 Liter.

The presence of Cu+2, Zn+2, and Mn+2 ions interfere with determination of hardness by this method.
Copper and zinc ions are removed by addition of 1ml of 1.5-2% Na2S solution. If Mn+2 is present
5 drops of a 1% solutions of hydroxylamine hydrochloride (NH2OH.HCl) is added before titration.
This is done to prevent oxidation of Mn+2 ions by atmospheric oxygen as it is the oxidation
products which interfere with the subsequent determination of hardness.

Procedure:

1) Take 25 ml of hard water in conical titration flask.

2) Add a few mg of Murexide Indicator into it.
3) Buffer Solution of PH 10 is also added in a quantity of 5-6ml.
4) Titrate this solution against standard EDTA solution taken in burette while stirring the
flask.
5) The end point is pink color is changed to violet.
6) Then note the reading.
7) In the same way, other readings will be taken.

Data Analysis:

Volume of water taken = X = 25 ml

Page | 24
Volume of EDTA used = Y = _______________ml
0.01M EDTA Solution = 0.01M CaCO3 Solution
. ×
1 ml of 0.01M EDTA solution = (CaCO3 in titrated volume of solution)

Then,
Calcium Hardness in terms of mg/liter or ppm of CaCO3
0.01 × 100 × 1000000
= × = 1000
1000

Page | 25
 Experiment 3
(ANALYSIS OF TOTAL HARDNESS OF WATER)

Objective:

Estimation of total hardness in water sample.

Equipment Setup / Apparatus:

Conical flask, Burette, Pipette, Iron stand, Oven.

Page | 26
 Oven

Reagents:

Hard Water, EDTA Solution, EBT Indicator, Buffer Solution.

Theory:

1. BUFFER SOLUTION
A buffer solution is a weak acid and a salt of that acid with strong base. The most favorable
buffer solution for this titration is prepared by using the mixture of NH4Cl/NH4OH having
PH=10. For this buffer solution take 70g of NH4Cl and add 568ml of conc. NH3OH
Solution and then stir and dilute the solution to 1 Liter with deionized water.
2. EBT
The indicator Eriochrome Black-T is used either in solution or in solid form (solution is
unstable). In the former case dissolve 0.2g of the pure solid dyestuff in 15ml of tri-ethanol

Page | 27
 amine. Add 5 ml of absolute ethanol to reduce viscosity. In the latter case, the indicator is
mixed with some filler such as NaCl or KCl in the Ratio 1:200. The mixture is ground in a
mortar and about 20 to 30 mg is added to the solution before titration.
3. EDTA / Standard Solution / 0.01 M
EDTA is the Di-Sodium Salt of Ethylene Diamine Tetra Acetic Acid. Dry 5 g of Di-Sodium
Di-Hydrogen ethylene diamine tetra acetic acid (Na2H2C10H12O8N2.2H2O) in oven at 80oC
for 1.5 hr. From dried salt take 3.7224g in water, dissolve it and make up to 1 Liter.

The presence of Cu+2, Zn+2, and Mn+2 ions interfere with determination of hardness by this method.
Copper and zinc ions are removed by addition of 1ml of 1.5-2% Na2S solution. If Mn+2 is present
5 drops of a 1% solutions of hydroxylamine hydrochloride (NH2OH.HCl) is added before titration.
This is done to prevent oxidation of Mn+2 ions by atmospheric oxygen as it is the oxidation
products which interfere with the subsequent determination of hardness.

Procedure:

1) Take 25 ml of hard water in conical titration flask.

2) Add a small amount of EBT Indicator into it.
3) Buffer Solution of PH 10 is also added in a quantity of 5-6ml.
4) Titrate this solution against standard EDTA solution taken in burette while stirring the
flask.
5) The end point is developed when violet red color is changed into dark blue color.
6) Then note the reading.
7) In the same way, other readings will be taken.

Data Analysis:

Volume of hard water taken = X = 25 ml

Volume of EDTA used = Y = _______________ml
0.01M EDTA Solution = 0.01M CaCO3 Solution
25 ml of H2O required 0.01M EDTA = ______________ml
25 ml of H2O required 0.01M CaCO3 = ______________ml
1 ml of H2O required 0.01M CaCO3 = ______________ml
1000 ml of H2O required 0.01M CaCO3 = ______________mg/L or ppm

Page | 28
Total Hardness in terms of CaCO3 = ______________ ppm of CaCO3
Calcium Hardness = ____________ ppm form Experiment-2
Magnesium Hardness of Water in terms of CaCO3 = Total Hardness – Calcium Hardness

Page | 29
 Experiment 4
(ANALYSIS OF SUGAR IN MILK)

Objective:

Estimation of Lactose in Milk.

Equipment Setup / Apparatus:

Volumetric Analysis Apparatus (Conical Flask, Pipette, Burette, Iron Stand, Wire Gauze
Tripod Stand, Burner, Beakers, Funnel, Whatman Filter Paper No. 41-42.)

Page | 30
Reagents:

Glucose, CuSO4.5H2O, Distilled Water, Rochelle salt (Sodium Potassium Tartrate), NaOH
or KOH, Methylene Blue Indicator, Milk Sample, Diluted Acetic Acid.

Theory:

Milk is the secretion of lectical glands of female mammals for the nourishment of child. It
contains all the requisites for a complete food i.e. sugar, fat, protein, mineral ingredients in
appropriate proportions. It is a pale, yellow or white opaque fluid denser than water. It is complete
solution of the sugar, soluble albumin, and mineral content, and in less complete casein while the
fat globules are held in suspension in the serum, forming an emulsion. The various mammals have
the same general physical properties and the same ingredients different in percentiles and the
animal’s milk is acidic while the human milk is alkaline to methyl orange but amorphous in nature.
The acidity of milk increases by keeping it because of lactic acid formation. The colors depend
upon seasons and other contents. Fat is the most variable constituent of milk ranging to nearly 7%,
lactose from 3 to 6%, protein (casein) about 3%, citric acid about 0.1%, mixture of water, fat
carbohydrates, proteins, and mineral matter together with chemical compounds. Organic
compounds may be cholesterol, carotene, lactochrome, globules enzymes and citric acid, gases are
CO2, O2, N2, and Mineral constituents are potassium, sodium, magnesium, and trace of iron,
copper, and zinc. The non-metallic elements include chlorine, sulfur and iodine.

For chemical analysis of milk, the usual determinations, in ascertaining the nutrition of
milk are:

1) Specific Gravity
2) Total Solids
3) Ash
4) Fat
5) Protein
6) Lactose

The specific gravity of pure milk at 60oF generally ranges from 1.027~1.036. Composition is
approximately as follows:

Page | 31
 1) Butter Fat 3.6%
2) Solids not fat 9.1%, casein 3%, other nitrogenous substances 0.8%, lactose 4.5%, ash 0.7%,
others 0.1%.
3) Total solids 3.6%+9.1%=12.7%

HYDROLYSIS REACTION

→ +

This is a general method to estimate sugar, glucose or any water-soluble carbohydrate in a

material. Lactose estimation can be performed by:

1) Fehling’s Solution Method

2) Polarimetric Method
3) High Performance Liquid Chromatography HPLC
4) Iodometric Method

Glucose Solution

Prepare 0.5% glucose solution by dissolving 0.5g of glucose in 100 ml of water.

Fehling Solution

a) Fehling – A Solution can be prepared by dissolving 22.149g of CuSO4.5H2O in water

diluting to 500ml.
b) Fehling – B Solution can be prepared by dissolving 173g of Rochelle salt (sodium
potassium tartrate) and 50g of NaOH or 125g of KOH in water and diluting to 500ml.
Fehling solution is first standardized with 0.5% glucose solution. Take 0.5% glucose
solution in the burette and 1ml of Fehling – A Solution and 1ml of Fehling – B Solution in
the titration flask. Dilute the Fehling solution mixture with 30ml of water and boil by
heating the flask over a wire gauze. Run in the glucose solution to the boiling solution
adding 2ml at a time and stirring vigorously. When only a faint blue color remains, the
glucose solution should be added in smaller quantities keeping the mixture near the boiling
point. When blue color of the mixture is considerably diminished in intensity, add 4 to 5
drops of Methylene Blue indicator. The solution will again turn deep blue. Again, run in
glucose solution dropwise till a sudden change from blue to red color appears due to the

Page | 32
 formation of cuprous oxide (Cu2O). That is the end point. Note the no. of ml. of 0.5%
glucose solution used. So, the Fehling’s solution has been standardized. Let x ml of the
glucose solution is required during this standardization.

Procedure:

1) Take 20g of milk in a 100ml beaker and it is diluted with about 60ml of distilled water.
2) Then add 3-4ml of diluted acetic acid.
3) Heat it on wire gauze for some time.
4) The casein separated out carrying with it fat also.
5) The clear liquid was obtained by filtration using Whatman 41 or 42. This liquid contains
lactose.
6) The lactose solution was then taken in the burette to determine its amount by titrating
against 1ml Fehling – A solution + 1ml Fehling-B solution mixture exactly in the same
way as with 0.5% glucose solution.
7) The dilution of milk was done to obtain a solution of lactose which does not contain more
than 1% of the substance, as with more conc. solution, the method does not give accurate
results.

Data Analysis:

1ml of Fehling – A Solution + 1ml of Fehling – B Solution = 2ml = 0.0106g of glucose

(2.12ml of 0.5% glucose solution required)

Milk Taken = 20g

GLUCOSE SOLUTION
Initial Reading of Glucose = _________________ml
Final Reading of Glucose =__________________ml
Volume of Glucose used = X = _______________ml

LACTOSE SOLUTION (FILTRATE)

Initial Reading of Filtrate = _________________ml

Page | 33
Final Reading of Filtrate =__________________ml
Volume of Filtrate Used = Y = ______________ml

Volume (ml) of milk extract required to react with 2ml (1ml A + 1ml B) of Fehling solution
mixture = Y
It is obvious that glucose volume X = lactose %
The amount of glucose in X ml = (0.5X)/100 g
The lactose content in g in y ml milk extract = (0.5X)/100 g
.
The lactose content in 100 ml milk extract =

% = × 100

0.5
% = × 100 = 2.5 ×
20

Page | 34
 Experiment 5
(ANALYSIS OF FAT IN MILK)

Objective:

Estimation of Fat Full Casein in Milk.

Equipment Setup / Apparatus:

Gravimetric Analysis Apparatus (Weight Balance, Whatman Filter Paper No 41 or 42,

Funnel, Iron Stand, Beakers, Pipette, Tripod Stand, Wire Gauze, Burner, Oven)

Page | 35
Oven

Page | 36
Reagents:

Distilled Water, Milk Sample, Diluted Acetic Acid.

Procedure:

1) Take 20g of milk in a 100ml beaker and it is diluted with about 60ml of distilled water.
2) Then add 3-4ml of diluted acetic acid.
3) Heat it on wire gauze for some time.
4) The casein separated out carrying with it fat also.
5) The clear liquid was obtained by filtration using Whatman 41 or 42. This liquid contains
lactose. And the residue on the filter paper contains fat full casein.
6) Dry this casein on the filter paper using oven to a constant weight. And then use gravimetric
analysis to determine Fat full casein percentage.

Data Analysis:

Weight of Milk Sample = 20 g

Weight of Filter Paper = X = ______________ g
Weight of Filter Paper + Residue = Y = _______________ g
Weight of Dried Residue = Z = Y-X = ________________ g

% = × 100

% = × 100
20

Page | 37
 Experiment 6
(ANALYSIS OF LEATHER)

Objective:

Determination of ash content of leather.

Equipment Setup / Apparatus:

Gravimetric Analysis Apparatus (Weight Balance), Crucible with Lid, Burner, Iron Stand,
Holder, Muffle Furnace, Desiccator.

Page | 38
 Furnace

Reagents:

Sample of Leather

Theory:

Leather, a common place item of everyday life, is the product obtained from animal skins
by several methods. From ages, it has been used for clothing, floor covering protection. Now a
days leather has got very much importance as a building material, shoes, bags, jackets, belts etc.

Page | 39
 Animal skin is made up of cells and fibers with some amorphous material the chemistry of
skin is concerned, it is made up of cells and fibers with some amorphous material the chemistry of
skin is concerned, it is made up chiefly of protein, but like all others contains lipids, carbohydrates,
inorganic salts and water. The proteins of the skin and components from a leather making
standpoint. They include collagen (which is a fibrous protein of the skin), Elastin, Reticulin,
Myosin (The muscle protein), albumin, globulin, and mucoid (non-flow fibrous proteins, which
do not make leather, but their complete absence will affect the quality of the finished leather).

The chemical analysis of vegetable tanned leather includes the following determinations:

1) Moisture
2) Ash
3) Fat and Oil
4) Water Soluble Matter (Total Water Soluble, Soluble Ash, Glucose)
5) Hide Substance
6) Combined Ash
7) Free Mineral Acid
8) Insoluble Ash
9) Degree of Tannage

Vegetable tanned leather may be considered as a mixture of the hide substance tanning
complex, with variable quantities of moisture, fatty matter, mineral matter and water soluble
matters e.g. unbounded tanning material. Sample of leather should be in fine state of division as
practicable, either by cutting with a sharp blade or grinding in wily will or rasping. Ash of leather
is composed of oxides of inorganic metals. The usual adulterants are barium sulphate and
magnesium sulphate. The ash of a good tanned leather should not exceed 2%.

Procedure:

1) 2g of the given sample of leather was taken in a prepared (cleaned by heating) and weighted
crucible.
2) The leather was charred (burnt) with lid on the crucible, in the open air, on the burner.
3) Strong heating was continued after removing the lid.

Page | 40
 4) And then the crucible containing charred sample of leather was placed in the muffle furnace
maintained at 1000oC and it was ignited to a constant weight.
5) Then it was cooled to room temperature and weighed again.
6) Then the percentage of ash in the sample of leather was calculated.

Data Analysis:

Weight of Crucible = X = ___________ g

Weight of Crucible with Leather = Y = ______________ g
Weight of Crucible with Ash = Z = ____________ g
Weight of Leather = A =Y-X = ___________ g
Weight of Ash = B = Z-X = ____________ g

% = × 100

Page | 41
 Experiment 7
(ANALYSIS OF VEGETABLE OIL)

Objective:

Determination of saponification value and saponification equivalent of a vegetable oil.

Equipment Setup / Apparatus:

Volumetric Analysis Apparatus (Weight Balance, Tripod Stand, Beakers, Burner, Wire
Gauze, Iron Stand, Burette, Pipette, Conical Flask, Spherical Bottom Flask, Condenser of Flask,
Water Bath).

Page | 42
Reagents:

KOH, HCl, Phenolphthalein Indicator, Vegetable Oil Sample, Distilled Water, Ethanol
(Alcohol 95% V/V).

Theory:

The saponification value is defined as the no. of milligrams of potassium hydroxide which
are required to saponify completely one gram of an oil, fat or wax.

The saponification equivalent is the number of grams of the substance saponified by

56.1mg (i.e. one equivalent of KOH in mg) of KOH.

A known weight of the oil is heated with a measured volume of a standard solution of
alcoholic potassium hydroxide, until saponification is complete. The excess of KOH remaining in
the solution is determined by titration with a standard solution of an acid (usually HCl) with
phenolphthalein as an indicator. The amount of KOH consumed for saponified amount of KOH
solution will be used for calculating the saponification value and saponification equivalent.

For evaluating the quality of a fat or oil, the tests performed are:

1) Moisture and Volatile Matter

2) Refractive Index
3) Saponification Value and Saponification Equivalent
4) Meissl and Polenske Values
5) Kirrchner Value
6) Acid Value
7) Mineral Acidity
8) Ester Value
9) Unsaponifiable Matter
10) Sterol Acetate Test
11) Hehner Valve

REAGENTS:

1) 0.5N HCl, Accurately Standardized

Page | 43
 2) 0.5N Alcoholic KOH Solution
It is prepared by dissolving 35 to 40 g of KOH pellets in 20 ml of water, mixing the solution
with one litter of alcohol 95% V/V, allowing the mixture to stand and pouring or filtering
off the clear supernatant liquid. Keep solution in glass stoppered bottle.
3) Phenolphthalein Indicator
1% in alcohol (95% V/V).

Procedure:

1) 8.197g of sample was taken in a 250ml Spherical Bottom Flask and 40 ml of the alcoholic
KOH were added by pipette.
2) The flask was connected with condenser and was heated on a boiling water bath so that the
alcoholic solution would boil gently for 30 mints.
3) When saponification was completed after 30 min and the solution appeared to be clear, the
flask was removed from the bath, cooled a little and was made to known volume with
water.
4) Excess alkali was titrated with acid by adding 1ml of indicator.
5) Three concordant readings were taken.

Data Analysis:

A = No. of ml of 0.5N acid required in the blank test.

B = No. of ml of 0.5N acid required in the sample test
W = Weight of oil = _______________ g
Volume of 0.5N KOH Alcoholic used = 40 ml
Solution taken for each titration = 10 ml
Solution in the burette = 0.5N HCl
Sr. No. Initial Reading (ml) Final Reading (ml) Difference (ml)

Average Volume used = V = ___________ ml

Volume of made up solution = A = ______________ml

Page | 44
Volume of 0.5N HCl required for X ml of Other Solution = B = (V/10)*100 =_______ ml
Volume of 0.5N HCl in terms of 0.5N Alcoholic Solution = B = __________ ml
Volume of 0.5N alkali used for saponification = A-B = _____________ ml

= × 28.05

56.1
=

Page | 45
 Experiment 8
(ANALYSIS OF SOAP)

Objective:

Estimation of fatty matter in soap.

Equipment Setup / Apparatus:

Beaker, Glass Rod, Water Bath, Volumetric Analysis Apparatus, Desiccator, Weight
Balance.

Page | 46
Reagents:

Soap Sample, Distilled Water, HCl, Stearic Acid, NaOH, Methyl Orange Indicator,
Phenolphthalein Indicator, Ethanol.

Theory:

Soap is a salt of sodium, potassium and ammonium of the non-volatile fatty acids (saturated
and unsaturated, beginning with those containing eight and higher carbon atoms). Strictly
speaking, all metallic salts of fatty acids are soap, but the salts of the alkaline metals and of
ammonium are appreciable water soluble used for cleansing. Some organic-base salts are also
soluble in water and are called detergents. The soaps of other metals e.g. calcium, aluminum,
magnesium, zinc, lead etc. are all insoluble in water and are useless for detergent purposes but they
are used for many other purposes such as for greases, water-proofing, color-printing and dyeing,
paints and varnishes, driers, plasters, plasticizers, fungicides, flatting agents, glazing agents. Three
processes are employed for soap manufacture namely when glycerol is not set free, glycerol is set
free but not separated and when no glycerol is formed. There are various kinds of soap e.g. milled
or toilet soap, transparent soap, textile soap, castile soap, mottled soap, shaving soap, medicated
soap, formaline soap, floating soap, liquid soap and dry cleaning soap.

Great number of substances are incorporated into soaps e.g. rosin, sodium, silicate and
borax, petroleum naphtha, coloring matter and perfumes. Scouring soaps often contain sand,
kiselgular, powdered and pumice. Some soaps contain flur, Sulphur, talc, china clay, salt etc.
Persulphates, per carbonates and perforates give bleaching and disinfecting properties.

Analysis of soap include the determination of combined fat, free fat, rosin, free caustic
alkali, carbonated alkali, combined alkali, water, sodium silicate, sodium chloride, sodium sulfate,
borax, mineral fillings, sugar, glycerin.

The estimation of the combined fat or uncombined alkali are important to find
thoroughness of saponification. Number of determinations are to be decided on the basis of kind
of soap.

Fatty matter may be present in free or combined form. Free fatty matter estimation is
performed by titrating oil or soap with standard alkali. For estimation of combined fatty acid, the

Page | 47
method includes the liberation of the fatty acid with the help of strong mineral acid. The acid so
formed is then estimated by weighing or by titrating with standard alkali.

To make the standard analysis of all the material, organic or inorganic, the analysis on the
dry basis should be performed to get reproducible results.

Procedure (For Total Fatty Acids):

1) This include soluble and insoluble fatty acids.

2) 5g of soap were dissolved in about 100ml of hot water in a beaker.
3) 40ml of 0.5N HCl were added. The beaker was heated on steam bath, until the insoluble
fatty acids were collected in a clean layer at the top of the liquid.
4) 10g of stearic acid were added to the hot liquid to effect it with above clear layer.
5) The beaker was cooled by placing in cold water and a cake formed. The removal of the
cake was facilitated by placing a glass rod with its fattened end into the beaker.
6) The solid cake was carefully removed from the beaker and was washed with distilled water.
7) The excess of water was removed by pressing the cake between the folds of filter paper. It
was dried in desiccator and was weighed.
8) This gave the weight of total fatty acids and the fats added.
9) To get the total fatty acids present in 5g of the sample, the weight of fatty acids added was
subtracted.
10) Calculate the percentage which may be multiplied by 0.97 (0.97673) to give the percentage
of fatty acid anhydrides of insoluble fatty acids.
11) The liquid after the removal of cake was titrated as it contains excess of 0.5N HCl, with
0.1N NaOH using methyl orange as indicator. Calculate the amount of 0.5N Acid
consumed in terms of Na2O, or K2O to find total alkali.
12) Now it was again titrated with 0.1N NaOH by using phenolphthalein as indicator till
reddish pink color appeared.
13) The amount of alkali used measure the amount of soluble fatty acid which were calculated
as caprylic acid.
1ml of 0.1N NaOH = 0.0144g Caprylic Acid

Procedure (For Free Fatty Acids):

Page | 48
 1) Dissolve 5g of soap in 95% Hot Alcohol (neutralized to phenolphthalein with 0.1N NaOH)
using hot water funnel, wash with hot neutral alcohol the residue of filtration.
2) The filtrate is titrated with standard alkali using phenolphthalein indicator.
3) Similar method is used to estimate free alkali.
1ml of 0.1N Acid (or Alkali) = 0.0040g NaOH
1ml of 0.1N Acid (or Alkali) = 0.0056g KOH
1ml of 0.1N Acid (or Alkali) = 0.0282g oleic acid
Data Analysis:

Reaction: C17H35COONa + HCl  NaCl + C17H35COOH (Caprylic Acid)

Solution in the Burette: 0.1N NaOH
Indicator: Methyl Orange
End Point: Orange Yellow
Indicator: Phenolphthalein
End Point: Reddish Pink
Weight of Sample = W = 5g
Volume of soap and water taken = 100ml
0.1N HCl added = 40ml
Volume of filtrate made = 150 ml
Volume of filtrate taken for each titration = 25ml
Volume of 0.1N NaOH for 10 ml sample = X = _______________ ml
Volume of 0.1N NaOH used for 250ml sample = (X/10) × 250 = ______________ ml
1ml of 0.1 NaOH = 0.0144g caprylic acid
25 X ml NaOH = 25 X*0.014g caprylic acid
Weight of Cake = M = _______________ g
Weight of Stearic Acid added = 10g
Sr. No. Indicator Initial Reading (ml) Final Reading (ml) Difference (ml)
1 Methyl Orange
2 Methyl Orange
3 Methyl Orange
Average

Page | 49
1 Phenolphthalein
2 Phenolphthalein
3 Phenolphthalein
Average

× 250 × 0.0144 × 100

% = = %
10 × 5

10
% = × 100 = %
5
% = % + % = ___________________

Page | 50
 Experiment 9
(ANALYSIS OF SILICATE)

Objective:

Estimation of silica (SiO2) in Sodium Silicate (Na2SiO3).

Equipment Setup / Apparatus:

Glass rod, Iron Stand, Tripod Stand, Burner.

Page | 51
Reagents:

Na2SiO3, Distilled Water, HCl, Silica gel.

Theory:

Silica is present in every rock. Analysis of rock requires that its solution be prepared. There
are certain products also which are silicates in nature and then aqueous solution is also made.
Various methods are used for making these solutions. These are dissolving in water and
decomposition by acid, fusion with alkali or reaction with hydrofluoric acid. Limestone and
cement decomposed with acids. Sodium silicate may be dissolved in water. Ceramic glass and
rocks need to be fused with fluxes such as sodium carbonate, sodium carbonate with calcium
carbonate, sodium carbonate with potassium nitrate, caustic soda and so on. Ceramic, glass and
rocks are reacted. These methods break the bonds and equilibrium is shattered which causes
solution in a liquid. Once the solution has formed, the remaining method is same in all the cases.

The solution obtained above in an acid medium, HCl is usually used, is made and it should
be completely transparent solution. No particle should be present. If some particles are present,
then its solution is not acceptable for analysis. The solution must be particle free. When such a
solution has been obtained, it has to be evaporated till a thick paste is obtained. Sand bath can be
used for this purpose. The paste should be dried on a water bath or steam bath till whole mass gets
dried completed. Further heating is continued till there is not smell of the acid. If whole drying
operation is done on a low temperature, then better results are obtained but sufficiently long time
is required for that. This operation is called baking. The baked mass must not attain red color. It
may be whitish or yellowish in color. The purpose of baking is to convert silicic acid (SiO2.H2O)
to SiO2 by dehydration. The solution obtained as a result of baking is soluble in dilute HCl solution
but silica (SiO2) does not dissolve and can be filtered out. The residue obtained on the filter paper
is washed thoroughly, dried and ignited at about 1000oC to a constant weight. For the estimation
of silica only gravimetric method is used.

Procedure:

1) 1 to 2g of sodium silicate paste was dissolved in 50ml of H2O by boiling 50ml of 0.5N HCl
solution were added and digested for half an hour.

Page | 52
 2) Then evaporated and baked to get a dry material free of HCl smell.
3) The solid mass was dissolved in 100ml dil. HCl and the residue was filtered out by using
the weighed filter paper.
4) The residue with filter paper was dried to a constant weight.
5) Then silica percentage was calculated.

Data Analysis:

Weight of silica sample = W = _______________ g

Solution of silica sample used = ______________ ml
0.5N HCl added = 50ml
Volume of conc. HCl added = 10ml
Volume of distilled H2O added = 10ml
Weight of filter paper = W1 = ______________ g
Weight of filter paper with residue = W2 = ______________ g

% = × 100

% = × 100

Page | 53
 Experiment 10
(ANALYSIS OF CHROMATE)

Objective:

Estimation of chromium in potassium chromate (K2CrO4).

Equipment Setup / Apparatus:

Volumetric analysis apparatus (Beakers, Tripod Stand, Wire Gauze, Burner, Test Tubes,
Iron stand, Flask, Pipette, Burette)

Page | 54
Reagents:
K2CrO4, Distilled Water, AgNO3, Ammonium per sulphate (NH4)2S2O8, KMnO4, O-
Phenanthroline Indicator, Mohr’s Solution.

Procedure:

1) Take 0.1g of K2CrO4 in a 400ml beaker and dissolved in 40ml of distilled water.
2) The solution was heated for about 10 to 15 minutes, cooled, 100ml of water were added
and then boiled for 3 minutes.
3) The solution was diluted to 200ml with water 15ml of AgNO3 solution, 3g of (NH4)2S2O8
(Ammonium per Sulphate) and a few drops of KMnO4 solution were added to the warm
solution to oxidize chromium and a pink color was developed for 5 minutes on boiling.
4) It was boiled for 5-10 minutes to remove HCl and Cl2. It was cooled and diluted to 300ml.
5) 10 ml of solution was taken for each time and a few drops of O-Phenanthroline indicator
were added.
6) It was titrated with Mohr’s solution till color changed and an excess known volume of
Mohr’s solution was added.
7) Excess volume was back titrated with standard KMnO4 solution. The consumed volume
will tell the amount of chromium.

Data Analysis:

Weight of sample (K2CrO4) = ________________ g

Mohr’s solution used = 0.3ml

Excess added = V = 5ml
Normality of Mohr’s solution = N1 = 0.1N
Normality of KMnO4 solution = N2 = 0.03N
KMnO4 solution used = V2 = ______________ ml
Excess of Mohr’s solution: N1V1 = N2V2
V1 = ___________ ml
Mohr’s solution added = V3 = V+V1 = ____________ ml
Consumed = V3-V1 =_______________ ml
Factor = 300/10 = 30

Page | 55
1ml of 0.1N Mohr’s Solution = 0.001733g Cr
0.05ml of 0.1N Mohr’s Solution = 0.00008665g Cr
.
% of Cr = × 100 × 30 = _______________ %
.

Page | 56
 Experiment 11
(ANALYSIS OF NaOH SOLUTION)

Objective:

Standardization of NaOH using standard HCl solution via back titration.

Equipment Setup / Apparatus:

Glass Wares (Petri Dish, Conical Flash, Burette, Pipette, Funnel, Crucible with Lid,
Spherical Bottom Flask, Condenser of Flask, Glass Rod, Test Tube).

Page | 57
Reagents:

Distilled Water, Standard HCl Solution, Standard Na2CO3 Solution, NaOH Solution of
Unknown Concentration and Methyl Orange Indicator.
Procedure:

1) Prepare standard solution of HCl (0.02M) and Na2CO3 (0.01M).

2) NaOH solution of unknown concentration is provided.
3) Take 10ml of NaOH solution in conical flask and add excess (20ml) but known amount of
4) Standard HCl solution in it.
5) Add one drop of methyl orange indicator.
6) Titrate it against standard Na2CO3 solution till indicator changes its color.
7) Calculate the amount of free acid left after neutralization with NaOH solution.
8) Calculate amount of acid neutralized by NaOH.
9) Calculate the concentration of NaOH solution.

Data Analysis:

+2 →2 + +
Na2CO3: HCl

M1V1:

V2 (Amount of free acid left after neutralization with NaOH) =

ν (Acid used by NaOH) =Total Acid added-V2

NaOH: HCl

M1V1 = M2ν

M1 (Concentration of NaOH unknown sample) =

Preparation of primary solution of HCl:

Molarity of 37% HCl can be calculated as

Page | 58
 370 ml pure HCl 1.19*1000 g pure HCl 1 gmole pure HCl
1000 ml of Primary HCl solution 1000 ml pure HCl 36.5 g pure HCl
Molarity of 37% solution= 12.06 M

Preparation of 250 ml of (0.02M) HCl from stock solution:

M1V1 = M2V2
( . ) ( )
V1 =
.
V1 = 0.414 ml 1000 µL
1ml
V1= 414 µL

Preparation of 250 ml of (0.01 M) Na2CO3 standard solution:

Number of moles of Na2CO3 needed = 0.01 *0.250 L

= 0.0025 mol
Mass of Na2CO3 = 0.0025*106
= 0.265 g

Preparation of 1000 ml (0.02 M) NaOH standard solution:

Number of moles of NaOH = 0.02 *1

Mass of NaOH = 0.02*40

Mass of NaOH = 0.8 g

Page | 59
 Experiment 12
(ANALYSIS OF BIOMASS)

Objective:

Estimation of moisture, volatile matter, fixed carbon and ash content in given sample of
biomass (Rice Husk).
Equipment Setup / Apparatus:

Furnace, Oven, Weigh Balance and Ceramic or Porcelain Crucible with Lid.

Oven and Furnace

Page | 60
 Furance

Procedure:

(i) Moisture Contents

Page | 61
 1) Took a clean, dry and properly weighed crucible. Note the weight after drying and recorded
it as initial weight of crucible.
2) Placed 1 gram of given sample in crucible and placed it in oven at 103˚C for 20 minutes.
3) After the 20 minutes removed the crucible from the oven and note down its weight and
again placed the sample in oven for 5 minutes.
4) Again removed crucible form oven after 5 minutes and note down its weight and placed it
again in oven for 5 minutes.
5) Repeated above step again and again till constant weight is achieved.
6) Note the weight that it was taken as initial weight for volatile matter.
(ii) Volatile Matter

1) Heated the crucible on burner with lid on it for 7 minutes.

2) Noted down its weight and recorded it.
3) The weight difference gave us weight of volatile matter.
4) Noted the weight and recorded it as initial weigh for ash.
(iii) Ash

1) Placed the crucible in furnace at 950˚C for 20 minutes. Removed it from the furnace and
noted down its weight again. Now again placed it into the furnace for 5 minutes.
2) Removed crucible again form furnace and noted down its weight.
3) Repeated above steps until constant weight is achieved. Noted down the weight.
4) Summed up moisture content, volatile matter and ash and subtracted it from total initial
value of to determine total fixed carbon.

Precautions:

1) As there is a lot of heat content so keep your lab coat, gloves and safety goggles on your
in order to avoid any injury.
2) Use clipper to hold apparatus and remain active throughout the practical.

Data Analysis:

(i) Moisture Contents

Page | 62
Initial weight of crucible =A=____________g
Weight of sample =B=____________g
Final weight =C=____________g
Weight of remaining sample =D= C-A=____________g
% Moisture Content = × 100

(ii) Volatile Matter

Final weight for volatile matter =E=____________g
Weight of remaining sample =F=E-A=____________g
% Volatile Matter = ×100

(iii)Ash
Final weight for ash =G=
Weight of remaining sample =H=G-A=____________g
% Ash content = × 100

Page | 63
SYNTHESIS BASED EXPERIMENTS

Page | 64
 Experiment 13
(SYNTHESIS OF PRUSSIAN BLUE)

Objective:

Synthesis of Prussian Blue.

Equipment Setup / Apparatus:

Beakers, Oven, Filter Paper, Funnel, Iron Stand.

Furnace and Apparatus

Page | 65
Reagents:

Ferric Chloride FeCl3.6H2O, Potassium Ferrocyanide K4Fe(CN)6.3H2O, Tap Water.

Theory:

Two blue color are usually used namely Ultramarine and Prussian blue. The iron salt, in
the ferric form, like ferric chloride, is reacted with ferrocyanide to precipitate ferric ferrocyanide
which is Prussian Blue. Mostly, ferric chloride is used but ferric sulphate can also be employed.
There are numerous routes for manufacturing ferric chemicals. Iron is reacted with any acid and
the iron salt is oxidized to convert ferrous to the ferric form.

Ferrous is prepared by reacting iron fillings, turnings or scrap with HCl. The solution so
obtained is then oxidized with the help of dilute nitric acid (any other oxidizing reagent may also
be used.) This ferric chloride solution is to be processed further in the synthesis of Prussian Blue.

Zinc chloride solution may be added to improve the color of the Prussian Blue as also to
increase the weight up to 1.25 times which will make the color cheap also.

Procedure:
1) Iron waste in the form of filling is reacted with HCl for making ferric chloride.
2) Ferrous chloride was then oxidized with nitric acid.
3) Ferrous chloride solution was heated to prepare a warm solution which was then removed
from the burner.
4) The solution container was kept in the glass or plastic trough to avoid sputtering during
boiling which would take place because of oxidation reaction concentrate nitric acid was
to be added in very small portions to fill a self-sustaining boiling.
5) 5g of Ferric Chloride was dissolved in the tap water.

6) 5g of Potassium Ferrocyanide was also dissolved in water.

7) Two solutions were mixed to precipitate the Prussian Blue color.

8) If precipitation did not occur, then added more volume of ferric chloride to effect
precipitation.

9) This precipitation took place in the acidic ranges.

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 Therefore, the residue was washed thoroughly with water.

1) The color so formed was oil-soluble and to make it water soluble solid potassium
ferrocyanide in the ratio of 1 to 7 (Cyanide to Prussian Blue on dry basis) was to be mixed.
2) The color was then dried in the oven.
3) This was a blue acidic pigment.

Reaction:

Fe + 2HCl → FeCl + ₂

1
2 +2 + →2 + ₂
2

1
2 +6 + →2 + 2 + +2 0
2

4FeCl + 3K [Fe(CN) ] → Fe [Fe(CN) ] + 12KCl

Salient Features:

1) This is acidic blue color and medium should be acidic during its applications.
2) Actually, ferric ion is required and this can be made from any water-soluble copper salt.
3) Oil-dispersible pigment is precipitated which is washed thoroughly. A very fine powder of
this material is required for use in paints and varnished.
4) By adding the ferrocyanide solid to the precipitated pigment in the ratio 1 to 7, the water-
soluble pigment forms.
5) Ferrous salt can be reacted with potassium ferrocyanide to prepare a similar blue acidic
pigment.

Synthesis Steps:

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 Experiment 14
(SYNTHESIS OF LIQUID SOAP)

Objective:

Synthesis of Liquid Soap.

Equipment Setup / Apparatus:

Beaker, Test Tube, Tripod Stand, Burner, Wire Gauze.

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Reagents:

Water, Caustic Potash KOH, Glycerin, Red Oil (Oleic Acid).

Theory:

Liquid soap to be used for washing purpose can be made by using potassium hydroxide,
oleic acid (usually called red oil which is unsaturated fatty acid). Shaving cream and sticks are
made by using potassium hydroxide by saponifying it with some fatty acid or oil.

Oleic acid or red oil is neutralized with caustic potash solution. The neutralized solution is
then heated to complete saponification and liquid soap of good quality is synthesized. Small
amounts of Sodium Silicate, Alcohol or Ether and urea may be added to the soap. Actually, after
washing with the liquid soap, the cloth should be rinsed in the solution of the above materials.
Some coloring and bleaching material may also be used in the preparation of the second solution.
Liquid soap solution may be named as washing solution whereas the other solution may be called
as the finishing solution.

Procedure:

1) A good grade liquid soap can be made by using the following recipe.
Water: 16g
Caustic Potash KOH: 1g
Glycerin: 4g
Red Oil (Oleic Acid): 4g
2) The caustic potash was dissolved in water and glycerin was added and brought to boil in
an enameled pot.
3) Then removed from heat and red oil was added slowly while stirring.
4) If more neutral soap was wanted, a little more quantity of red oil was used.

Reactions:

C H COOH + KOH → C H COOK + H O

(C H COO)C H (Oil) + 3KOH → 3C H COOK(Soap) + C H (OH) (Glycerin)

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Salient Features:

1) Usually solid soaps are preferred for washing purpose but commercial cleaning requires
liquid soap or liquid detergent. Solid material has to be dissolved in water for use.
2) Neutralization extent and the type of materials will decide whether the soap would be in
liquid or solid form.

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 Lab Flexes

1. Lab Notebook Rubric

2. Experiment Performance Rubric
3. List of Equipment
4. List of Experiments
5. IUPAC Periodic Table 2016
6. List of Atomic Weights

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