STK1211

Practical Analytical
Chemistry

Laboratory Manual

Department of Chemistry
Faculty of Resource Science & Technology
Universiti Malaysia Sarawak
2015/2016

Example of cover page

FACULTY OF RESOURCE SCIENCE AND TECHNOLOGY

DEPARTMENT OF CHEMISTRY

STK 1211 Practical Analytical Chemistry

EXPERIMENT NO:

TITLE OF EXPERIMENT:

DATE OF EXPERIMENT

GROUP MEMBERS & MATRIC

NUMBERS

LAB FACILITATOR

REPORT DUE DATE

Table of Contents
Page

Laboratory outline
-

Laboratory report format

Laboratory report submission

Group experiments

Laboratory safety

Apparatus and techniques

Clean and Clean and Dry glassware

Desiccator and handling dried compounds

Electronic and analytical balance

Volumetric flasks and quantitative transfers

Experiment 1: Acid-base Titrations

11-14

Experiment 2: Water Analysis: Suspended Solids and Dissolved Oxygen

15-19

Experiment 3: Water Analysis: Hardness of Water and Chlorine

Concentration

20-24

Experiment 4: Redox Titration Ascorbic Acid

25-29

Experiment 5: Gravimetric Determination of Nickel(II) ion

30-33

Experiment 6: Thin-layer Chromatography

34-38

Laboratory Outline
1. The lab report should be organized according to the following sequence:
a) Introduction
b) Objectives
c) Procedures and material/apparatus
d) Results
e) Calculations, if any
f) Discussion
g) Conclusion
h) Post-lab questions
i) References
2.
3.
4.
5.
6.
7.

Unorganized lab report will be penalized.

The content of each section in a lab report is described in Table 1.
Only one lab report needs to be submitted per group.
Lab report is to be submitted at the beginning of next lab session.
Late submission will be penalized.
Plagiarism is strictly prohibited.
Table 1: Description on the content of a lab report

Section
Introduction

Objectives
Experimental
set up and
procedure

Results

Calculation
(if any)

Description
Brief discussion on the background of the technique(s) used in the
experiment.
The theory behind any calculation involved should be presented in
the introduction as well.
Any referred information must be cited.
Brief description on the objectives of the outlined experiment
Consists of the list of chemicals and apparatus used.
All steps performed in the experimental procedure should be listed
in the order that they were performed, in exactly the manner in
which you performed them.
The procedures must be written in passive sentences.
Should list all data obtained, in raw form, with information
provided as to how the data was obtained, as well as the accuracy
of all measurements.
Record the numbers/measurements you collect (Quantitative data)
Record other pertinent observations (Qualitative data)
If possible use tables.
Include graphs if appropriate.
Include ALL, even those that will be rejected later.
Calculation should be included after the results shown.
Include formulas, units, use significant digits.
Show complete calculations for all results.

Discussion

The data should be discussed and evaluated, both positively and

negatively.
Do not try to manipulate data to fit the results you think you
should obtain.
Evaluate the data fairly, even if the data seem to contradict with
theory you may have been expecting the data to follow.
A discussion of possible sources of error should be included in this
section.
Calculate your percent error, if applicable.
If any data was rejected, explain why here.
Any referred information must be cited.
Include chemical reactions involved (if any).

Conclusion

Concise, direct statement of what you learned.

If possible, use a single sentence.
Answer all the post-lab questions provided after each experiment.

Post-lab
questions
References

All materials have been used in writing the laboratory report

should be listed (at least two).
Journal articles are referenced by listing the authors (last name
first), the year of publication, the title of the article, the title of the
journal, the volume number and the page number. For example:
Kuivinenm J., Johnsson, H. (1999) Determination of Trihalomethanes
and some Chlorinated solvents in Drinking Water. Water Research, 33
(5). 1201-1208.
Books are referenced by listing the authors (last name first), year
of publication, the title of the book, the edition, the publisher, city
of publication and the page number. For example:
John, M.M., Johnston, D.O., Nettervile, J.T., Wood, J.L., Joesten, M.D.
1991. Laboratory Manual to Accompany World of Chemistry. Sounders
College Publishing, New York.

Laboratory Safety
Safety in the laboratory is a subject of the utmost importance. All chemicals are harmful to
some degree, therefore it is imperative to learn the safety rules and follow them strictly at all
times. You will be expelled from the laboratory for failing to comply with these regulations.
These rules are referred to many laboratories as the usual safety procedures.
General
1.
2.
3.
4.
5.
6.
7.
8.

Wear shoes at all times when you are in the laboratory.

Wear lab coat at all times when you are in the laboratory
Report any spill or accident immediately to your instructor.
Know the location and operation of safety equipment in the laboratory from the first
meeting of the laboratory section.
Drinking, eating and smoking are absolutely forbidden in the laboratory.
Never work alone in the chemical laboratory.
Dispose the chemicals properly, in the container provided, and according to the
instructions given by the laboratory instructor. Do not simply pour waste chemicals
down the sink.
Keep your laboratory space clean.

Safety glasses
1. Safety glasses should be worn at all times while in the laboratory.
2. Contact lenses should never be worn in the laboratory because they cannot be
removed rapidly if reagents accidentally splash in the eye.
Chemicals
1. Handle all chemicals according to any specific directions indicated on the container, or
those given to you by your instructor.
2. Avoid contact with skin and clothing.
3. Wipe up spills immediately, especially near the balances and reagent shelf.
4. Replace caps on containers immediately after use.
5. Avoid the inhalation of organic vapours, particularly aromatic solvents and
chlorinated solvents.
Disposals of chemicals
1.
2.
3.
4.

Dispose of chemicals as directed in each experiment.

Water-soluble substances can be flushed down the drain with large quantity of water.
Water insoluble solids and liquid should be placed in the waste container provided.
Chromium ion in the +6 oxidation state should be reduced to the +3 state with a mild
reducing agent before disposal.

Apparatus and techniques

The following is a summary of the basic analytical laboratory techniques and equipment you
will use for this semester. Proper techniques are essential as acceptable error in a
quantitative chemical analysis is seldom greater than 0.1 %. There are a number of hard and
fast rules presented that must be followed to minimize any hazards to yourself, your labcoworkers, and the lab equipment. Read this section at the beginning of the semester
and refer to any of this material as often as necessary.
I. Clean and Clean and Dry glassware
You will notice throughout the semester that you are asked to use Clean glassware at times
and Clean and Dry glassware at other times. Clean glassware may be wet with your
solvent (usually distilled water), and many times it is not worth the effort to dry the piece of
glassware.
In general, if you want to maintain the concentration of the solution being transferred, you
will want the final container to be Clean and Dry. However, if you are only concerned about
the amount of the compound being transferred, the final container need only be Clean.
Clean glassware means that all compounds and materials have been washed out. The final
washing should be with your solvent. In analytical chemistry, the solvent is defined as the
liquid or solution that you would use to dilute the solution in question, usually distilled water.
II. Desiccators and Handling Dried Compounds
When using primary standards (compounds that are presumed to be 100% pure) in analyses,
it is essential that there are no crystal waters present so that the mass of the primary
standard measured on the balance is equal to the actual mass. Typically, compounds are dried
by placing them in an oven at 105 to 120 C. After several hours, all (or most) crystal waters
have been driven off. However hot compounds cannot be accurately weighed (all items
weighed on a balance must be at room temperature), so there must be a way to cool a
dried compound without re-exposing it to water vapor in the atmosphere.
Desiccators are containers designed to prevent the re-hydration of solids. The bottom half of
the desiccator is filled with an anhydrous salt, such as calcium chloride. The dried compound
and its container sit in the top half, which is separated from the bottom half by a grid or
screen. The desiccator lid can be sealed with vacuum grease to prevent water vapor from
seeping inside.
Always cool a dried compound in a desiccator before weighing. A dried compound can be kept
in the desiccator if that compound has to be available throughout the lab period.
III. Electronic and Analytical Balances
Electronic balances are quite simple to use. As you probably know, the "tare" button resets the
mass reading at zero, and there is usually another button (sometimes labeled "cal" for
calibrate) to set the mass units. You will always want the mass units in grams. Because
electronic balances are fragile, you need to observe the following guidelines.
1. Always clean the balance after using it -- use a soft brush or Kimwipe to remove any
extraneous material from the balance pan.
2. All items and compounds placed on the balance must be at room temperature -- this can
come into play when weighing dried compounds. Cool dried compounds in a desiccator
before weighing them on the balance.
3. If you are making repeated weighing in the same container, it is recommended that you
always tare the empty balance and record the mass of the empty container. Then, record
the mass of the container with sample, and calculate the mass of the sample by difference.

4. If you are instructed to "accurately weigh" something, use a balance with 4 decimal places.
This is referred to as analytical balance. The maximum capacity of an analytical balance
is usually small (60 or 180 g), therefore use only weighing boats or weighing paper as
containers on the balance. On the electronic balance with 3 decimal places, you can often
use small beakers or flasks as containers.
IV. Volumetric Flasks and Quantitative Transfers
Volumetric flasks are calibrated to contain an exact volume of solution when the solution level
is exactly at the mark on the neck of the flask (the bottom of the meniscus should lie exactly
at this mark). Note the following rules in handling volumetric flasks.
1. To clean volumetric flasks -- Each washing should have a volume that is about 10 to
20% of the capacity of the vol. flask. Typically, you should wash the flask 3 times with
dilute acid (e.g., 1 to 6 M HCl), 3 times with distilled water, and 3 times with your solvent
(if it is not distilled water). You can skip the acid washings if you have no solid residue in
the flask.
2. Never heat a volumetric flask -- heating causes the glass to expand, changing the
volume it contains.
3. NEVER place a solid directly into a volumetric flask -- What do you do if you fill the
flask to the mark and the solid won't dissolve? Dissolve the solid in another container and
quantitatively transfer the solution to the volumetric flask. For example, if you want to
make a 100-mL solution of NaCl, dissolve the NaCl in a beaker with 75 to 90 mL of water
and then transfer this to the volumetric flask.
4. Never shake a volumetric flask -- when mixing a solution in a vol. flask, gently invert
the flask 8 to 10 times.
5. Quantitative transfers are vital to accurate analyses. In simple terms, quantitatively
transferring something means washing the original container and all glassware involved
in the transfer with solvent and adding those washings to the final container, usually a
volumetric flask. Here are some guidelines to transferring solutions and solids to
volumetric flask.
Pour the solution through a funnel into the volumetric flask. Wash the original container with
a small amount of solvent and pour the washing through the funnel into the volumetric flask.
Repeat the washing 2 or 3 times if possible. Dilute the solution in the volumetric flask to the
mark.
V. Volumetric Pipettes
Volumetric pipettes are calibrated to deliver an exact volume of liquid or solution. Volumetric
pipettes have only one calibration mark. You may have seen graduated pipettes that have
calibration marks throughout the length of the pipette, but these are far less accurate
than volumetric pipettes. To fill a pipette, simply draw in liquid to the mark. Usually it is
easiest to initially overshoot the mark and then let the liquid drain from the pipette until the
bottom of the meniscus lies exactly on the calibration mark.

Note the following rules in handling pipettes.

1. Never pipette with your mouth! Always use a rubber bulb, regardless of what you are
taught in biology classes.
2. Always clean a pipette before its initial use -- For each washing, draw liquid into the
pipette so that the bulb is 1/4 to 1/2 full (this can be less for large pipettes). Carefully swirl
the liquid throughout the inside of the pipette (don't let liquid pour out the top!), and let
the liquid drain from the pipette. Wash the pipette 3 times with distilled water and 3
times with the solution you are going to transfer (not just the solvent). If you are using the
same pipette for different solutions, you need to repeat the washing procedure every time
you switch solutions.
3. Never force liquid out of a pipette -- always let the liquid drain of its own accord. The
calibration mark takes into account any liquid that is retained in the tip of the pipette.
When the liquid has stopped draining, touch the tip of the pipette against the side of the
container to release any hanging drops.
4. At the end of a lab period, always wash used pipettes 3 times with distilled water.
VI. Burettes
Burettes allow you to accurately deliver volumes of liquid that cannot be measured by
volumetric pipettes or micropipettors. The proper use of burettes is essential to accurate
titration analyses. In several cases, an analyte can be determined more accurately using a
titrimetric method rather than an instrumental method -- it's just that the convenience of the
instrumental use is sometimes the deciding factor in choosing an analytical method. Note the
following rules and guidelines in using burettes.
1. To clean a burette -- fill the burette with distilled water and drain a large portion of it to
see if any water adheres to the inside walls of the burette. If so, clean the burette with a
few milliliters of soap solution and a burette brush, and wash the burette with three 5-mL
portions of tap water. When washing the inside of a burette, pour about 5 mL of liquid
into the burette with the stopcock closed. Carefully swirl the liquid for a few seconds so
that it comes in contact with the entire inside surface area of the burette, and pour the
liquid out the top. If no water adheres to the inside walls of the burette, proceed to wash it
3 times with distilled water and 3 times with the solution with which you are going to
titrate your sample.

2. Getting rid of air bubbles -- fill the clean burette with your solution. There will be air
bubbles inside the stopcock, and you must remove these (you can't measure the volume of
an air bubble in a burette, so if an air bubble pops out in the middle of your titration,
you're sunk). It's usually easiest to force bubbles out the stopcock, so simply open the
stopcock and let the solution drain until the air bubbles are removed. If this isn't working,
you can try gently tapping the base of the burette while the solution is draining. This
may force the air bubbles to rise through the solution.
3. Always record volume levels to the nearest 0.01 mL -- Although the calibration marks
are only at every 0.1 mL, you can always estimate the extra decimal place.
For Example:
The reading in this picture is between 8.2 and 8.3 mL. The second decimal place would be
estimated to be about 0.07 mL, giving a reading of 8.27 mL.

To obtain readings, it helps to hold a white card behind the burette. Note reading at a
particular part of meniscus and always measure at this part.

4. NEVER record an initial volume level of 0.00 mL -- since there are no calibration
marks above 0.00 mL, you have no upper reference with which to base any possible error
in your reading.
5. Using your wash bottle, you can add "half-drops" to your titration flask (see above figure).
Run a drop of solution part-way out of your burette tip. Squirt distilled water on this halfdrop and into your titration flask. This procedure is perfectly legitimate because you
aren't worried about the concentration of anything in your titration flask.
6. At the end of an experiment, always wash your burette 3 times with distilled water.

10

EXPERIMENT 1: ACID-BASE TITRATIONS

INTRODUCTION
The technique of titration finds many applications, but is especially useful in the analysis
of acidic and basic substances. Titration involves measuring the exact volume of a solution
of known concentration that is required to react with a measured volume of a solution of
unknown concentration, or with a weighed sample of unknown solid. A solution of
accurately known concentration is called a standard solution.
In many cases it is possible to prepare a standard solution by accurate weighing of the
solute, followed by precise dilution to an exactly known volume in a volumetric flask. One
of the most common standard solutions used in acid-base titration analysis, however,
cannot be prepared in this manner.
Solutions of sodium hydroxide are commonly used in titration analysis of samples
containing an acidic solute. Although sodium hydroxide is a solid, it is not possible to
prepare standard sodium hydroxide solutions by mass. Solid sodium hydroxide is usually
of questionable purity. Sodium hydroxide reacts with carbon dioxide from the atmosphere
and is also capable of reacting with the glass of the container in which it is provided. For
these reasons, sodium hydroxide solutions are generally prepared to be approximately a
given concentration. They are then standardized by titration of a weighed sample of a
primary standard acidic substance. By measuring how many mL of the approximately
prepared sodium hydroxide are necessary to react completely with a weighed sample of a
known primary standard acidic substance, the concentration of the sodium hydroxide
solution can be calculated. Once prepared, however, the concentration of a sodium
hydroxide will change with time (for the same reasons stated above). As a consequence,
sodium hydroxide solutions must be used relatively quickly.
In titration analysis, there must be some means of knowing when enough titrant has been
added to react exactly and completely with the sample being titrated. In an acid-base
titration analysis, there should be a sudden change in pH when the reaction is complete.
For example, if the sample being titrated is an acid, then the titrant to be used will be
basic. When one excess drop of titrant is added, the solution being titrated will suddenly
become basic. There are various natural and synthetic dyes, called indicators, which exist
in different coloured forms at different pH values. A suitable indicator can be chosen that
will change colour at a pH value consistent with the point at which the titration reaction is
complete. The indicator to be used in this experiment is phenolphthalein, which is
colourless in acidic solutions, but changes to a pink form at basic pH.

APPARATUS
Burette with stand
Pipette
1-L plastic bottle with stopper
250 mL Erlenmeyer flasks
Retort stand with clamp

11

REAGENTS
Sodium hydroxide pellets
Potassium hydrogen phthalate (KHP)
Phenolphthalein
Unknown vinegar

PROCEDURE
Part A: Preparation of the Sodium Hydroxide Solution
1. Clean and rinse a 1-L volumetric flask and stopper. Label the flask Approx. 0.1 M
NaOH. Put about 500 mL of distilled water into the flask.
2. Weigh out approximately 4.0 g of sodium hydroxide pellets and transfer to the 1-L
flask. Stopper and shake the flask to dissolve the sodium hydroxide.
3. When all the sodium hydroxide pellets have dissolved, add additional distilled water
to the bottle until the mark on the neck of the flask. Stopper and shake thoroughly to
mix.
Part B: Standardization of the Sodium Hydroxide Solution
1. Set up the burette in the burette clamp. Rinse and fill the burette with the sodium
hydroxide solution just prepared.
2. Clean three 250-mL Erlenmeyer flasks with water, and then rinse with distilled water.
Label them as 1, 2, and 3
3. Remove the bottle of dried KHP from the oven. When the KHP is completely cool,
weigh three samples of KHP between 0.6 and 0.8 g , one for each of the Erlenmeyer
flasks. Record the exact weight of each KHP sample to the nearest mg ( 0.001 g).
4. Add 100 mL of distilled water to KHP sample 1. Add 2 3 drops of phenolphthalein
indicator solution. Swirl to dissolve the KHP sample completely.
5. Record the initial reading of the NaOH solution in the burette to the nearest 0.02 mL.
6. Add NaOH solution from the burette to the sample in the Erlenmeyer flask, swirling
the flask constantly during the addition.
7. When the titration approaching the endpoint, add NaOH one drop at a time, with
constant swirling, until one single drop of NaOH causes a permanent pale pink colour
that does not fade on swirling. Record the reading of the burette to the nearest 0.02
mL.
8. Repeat step 4 7 with the other 2 KHP samples.
9. Given that the molecular mass of KHP is 204.2, calculate the number of moles of KHP
in samples 1, 2, and 3.
10. From the number of moles of KHP present in each sample, and from the volume of
NaOH solution used to titrate the sample, calculate the molar concentration (M) of
NaOH in the titrant solution. The reaction between NaOH and KHP is of 1 : 1
stoichiometry.

12

Part C: Analysis of a Vinegar Solution

Vinegar is a dilute solution of acetic acid and can be effectively titrated with NaOH using
the phenolphthalein endpoint.
1. Clean three Erlenmeyer flasks, and label as samples 1, 2, and 3.
2. Rinse the 5-mL pipette with small portions of the vinegar solution and discard the
rinsings.
3. Using the pipetter, pipette 5 mL of the vinegar solution into each of the Erlenmeyer
flasks. Add about 100 mL of distilled water and 2 3 drops of phenolphthalein
indicator solution to each flask.
4. Refill the burette with the NaOH solution and record the initial reading of the burette
to the nearest 0.02 mL. Titrate Sample 1 of vinegar in the same manner as in the
standardization until one drop of NaOH causes the appearance of the pink colour.
5. Record the final reading of the burette to the nearest 0.02 mL.
6. Repeat the titration for the other two vinegar samples.
7. Based on the volume of vinegar sample taken, and on the volume and average
concentration of NaOH titrant used, calculate the molar concentration of the vinegar
solution.
8. Given that the formula mass of acetic acid is 60.0, and the density of the vinegar
solution is 1.01 g/mL, calculate the percent by mass of acetic acid in the vinegar
solution.

QUESTIONS
1. Give the definition of indicators.
2. Suppose a NaOH solution were to be standardized against pure solid primary standard
grade KHP. If 0.4538 g of KHP requires 44.12 mL of the NaOH to reach a
phenolphthalein endpoint, what is the molarity of the NaOH solution?
3. Commercial vinegar is generally 5.0 0.5% acetic acid by weight. Assuming this to be
the true value for your sample, by how much were you in error in your analysis?

13

Experiment 1: Acid and Base Titrations

Data Sheet
Name:
Student No.:
Date:
_______________________________________________________________________
Part A: Standardization of the Sodium Hydroxide Solution
Particulars

Trial 1

Trial 2

Trial 3

Mass of KHP taken (g)

Final burette reading (mL)
Initial burette reading (mL)
Volume of NaOH used (mL)
Molarity of NaOH solution
Average molarity of NaOH solution

Part B: Analysis of a Vinegar Solution

Particulars

Trial 1

Volume of vinegar solution used (mL)

Final burette reading (mL)
Initial burette reading (mL)
Volume of NaOH used (mL)
Molarity of NaOH solution
Molarity of vinegar solution
% mass of acetic acid in vinegar
Average molarity of vinegar solution
Average % mass of acetic acid in vinegar

14

Trial 2

Trial 3

EXPERIMENT 2: WATER ANALYSIS: SUSPENDED SOLIDS AND

DISSOLVED OXYGEN
INTRODUCTION
Water is a very important resource for human being. A large portion of our body is water.
A healthy human being can live for weeks without foods, but will not survive for days
without water. When you are thirsty and need a drink of water, do you take for granted
that the tap water is safe to drink? Do you think about what might be dissolved or
suspended in the water?
Water in the environment has a large number of impurities. Dissolved solids are watersoluble substances, usually salts. Naturally occurring dissolved solids generally result
from the movement of water over mineral deposits, such as limestone. These dissolved
solids generally consist of the sodium, calcium, magnesium, and potassium cations and the
chloride, sulfate, bicarbonate, carbonate, bromide, and fluoride anions. The dissolved
solids are responsible for the hard water that exists in some locales. Anthropogenic
(human-related) dissolved solids include nitrates from fertilizer runoff and human wastes,
phosphates from detergents and fertilizers, and organic compounds from pesticides and
industrial wastes. Salinity, a measure of the dissolved solids in a water sample, is defined
as the grams of dissolved solids per kilogram of water.
Suspended solids are very finely divided particles that are water insoluble but are
filterable. These particles are kept in suspension by the turbulent action of the moving
water. Examples of suspended solids include decayed organic matter, sand, salt, and clay.
Total solids are the sum of the dissolved and suspended solids in the water sample. In this
experiment the total solids and the dissolved solids are determined directly; the suspended
solids are assumed to be the difference, since total solids = dissolved solids + suspended
solids
The dissolved oxygen is determined based on the iodine/thiosulfate method. Excess
potassium iodide is added to the acidified water sample, the dissolved oxygen (or other
oxidizing agents that are present) will oxidize the iodide ions to iodine (in the form of
triiodide, I3 ). The iodine formed will then be determined by titration with the standard
solution of sodium thiosulfate with starch solution as the indicator.

APPARATUS
Burette with stand
Pipette
Beaker
250 mL Erlenmeyer flasks
Evaporation dishes
Watch glass
Filter funnel
Filter paper

REAGENTS

15

KI
0.05M Na2S2O3 solution
Starch solution
6M Sulfuric acid

PROCEDURE:
I: Determination of Suspended Solids in a Water Sample
1.

Clean, dry, and determine the mass (to the nearest milligram, 0.001 g) of two
evaporation dishes. Be certain that you can identify each. Use the same balance for
the remainder of the experiment.

Part IA: Determination of dissolved solids

1.

2.

3.

Gravity filter about 50 mL of a thoroughly stirred or shaken water sample into a

clean, dry 100 mL beaker. While waiting for the filtration to be completed, proceed to
Part B.
Pipette a 25 mL portion of the filtrate into one of the evaporating dishes. Determine
the mass of the combined evaporating dish and sample. Place the dish containing the
sample on the wire gauze and heat slowly (do not boil) the mixture to dryness. As the
mixture nears dryness, cover with a watch glass, and reduce the intensity of the
flame. If spattering does occur, allow the dish to cool to room temperature, rinse the
adhered solids from the watch glass and return the rinse to the dish.
Again heat slowly, and avoid further spattering. After all of the water has evaporated,
maintain a small flame beneath the dish for 3 minutes. Allow the dish to cool to room
temperature and determine its final mass.

Part IB: Determination of total solids and suspended solids

1.
2.

Thoroughly agitate 100 mL of sample; pipette a 25 mL aliquot of this sample into the
second evaporating dish. Evaporate the sample to dryness as described in Part A.
Calculate the mass of total and suspended solids in the original water sample, using
data from Part A.

Part IC: Analysis of data

1.
2.

Compare your data with three other groups in your laboratory who analyzed the same
water sample. Record their results on the data sheet.
Calculate the standard deviation of the suspended solids from the four analyses on the
water sample.

II: Determination of Dissolved Oxygen in a Water Sample

1.

Prepare three clean Erlenmeyer flasks.

2.

Clean a burette with tap water, then rinse with distilled water, and finally rinse with
the 0.05M sodium thiosulfate standard solution. Then fill the burette with the sodium
thiosulfate solution and record its initial volume to the nearest 0.02 mL

16

3.
4.

Using a clean pipette, transfer 25 mL of the water sample into each of the Erlenmeyer
flasks and label as sample 1, 2 and 3.
Add about 2 g of KI and 10 mL of 6M sulfuric acid to sample 1. Iodide ions will be
oxidized to iodine, and then it is titrated with the standard sodium thiosulfate
solution.

5.

On titration, the colour of sample will become lighter. Stop adding the Na2S2O3
solution when the colour of the sample becomes light yellow that means it has nearly
reached the end point. Now add 2 3 mL of starch solution to the sample. The
sample will appear in dark-blue colour.

6.

Continue adding the Na2S2O3 solution drop by drop until the dark-blue colour just
disappears. Record the final volume to the nearest 0.02 mL

7.

Repeat the titration with the sample 2 and 3. From the concentration and the volume
of the Na2S2O3 solution used in the titration, calculate the concentration of oxygen
present in the water sample.

QUESTIONS
1.

In evaporating a solution to dryness in an evaporating dish, why must the heating rate
be decreased as the mixture nears dryness?

2.

How do pollutants such as sewage lower the dissolved oxygen content of water
sources?

3.

A 25 mL aliquot of a well-shaken sample of river water is pipetted into 27.211 g

evaporating dish. After the mixture is heated to dryness, the dish and remaining
sample has a mass of 43.617 g. Determine the total solids in the sample, express in
units of g/kg sample. Assume the density of the sample to be 1.01 g/mL.

17

Experiment 2: Water Analysis: Suspended Solids and Dissolved Oxygen

Data Sheet
Name:
Student No.:
Date:
______________________________________________________________________________________

I: Determination of Suspended Solids in a Water Sample

Part IA: Determination of dissolved solids
Particular

Data

Mass of evaporating dish (g)

Mass of evaporating dish + water sample (g)
Mass of water sample (g)
Mass of evaporating dish + dried sample (g)
Mass of dissolved solids in 25 mL aliquot (g)
Mass of dissolved solids per total mass of sample (g
solids/g sample)
Dissolved solids (g solids/kg sample)

Part IB: Determination of total solids and suspended solids

Particular

Data

Mass of evaporating dish (g)

Mass of evaporating dish + water sample (g)
Mass of water sample (g)
Mass of evaporating dish + dried sample (g)
Mass of total solids in 25 mL aliquot (g)
Mass of total solids per total mass of sample (g
solids/g sample)
Suspended solids (g solids/kg sample)

18

Part IC: Analysis of data

Particular

Group 1

Group 2

Group 3

Group 4

Dissolved solids (g/kg)

Total solids (g/kg)
Suspended solids (g/kg)
Average Suspended solids (g/kg)

II: Determination of Dissolved Oxygen in a Water Sample

Concentration of Na2S2O3 standard solution: _______________________________
Particular

Trial 1

Final burette reading (mL)

Initial burette reading (mL)
Volume of S2O32 used (mL)
Moles of S2O32 used (mol)
Volume of water sample (mL)
Concentration of sample (M)
Mean concentration of sample (M)

19

Trial 2

Trial 3

EXPERIMENT 3: WATER ANALYSIS: HARDNESS OF WATER

AND CHLORIDE ION/CHLORINE ION
CONCENTRATION
INTRODUCTION
What does it mean when we say that water is hard? Hard water contains the dissolved
salts of calcium, magnesium, and iron ions which are called hardening ions. In low
concentrations these ions are not considered harmful for domestic use, but at higher
concentrations these ions interfere with the cleansing action of soaps by forming insoluble
compounds with soaps. Soaps, sodium salts of fatty acids such as sodium stearate,
C17H35CO2Na, are very effective cleansing agents so long as they remain soluble; the
presence of the hardening ions however causes the formation of a grey, insoluble soap
scum such as (C17H35CO2)2Ca. This grey precipitate appears as a bathtub ring and it also
clings to clothes, causing white clothes to appear grey.
In industries, hard water can accelerate the corrosion of steel pipes, especially those
carrying hot water. It is also responsible for the formation of boiler scale on tea kettles
and pots used for heating water. The boiler scale is a poor conductor of heat and thus
reduces the efficiency of transferring heat. Boiler scale also builds on the inside of hot
water pipes to decrease the flow of water; in extreme cases, this buildup causes the pipe to
break. Boiler scale consists primarily of the carbonate salts of the hardening ions. Ground
water becomes hard as it flows through underground limestone (CaCO3) deposits; Surface
water similarly accumulates hardening ions as a result of it flowing over limestone
deposits. Because of the relative large natural abundance of limestone deposits and other
calcium minerals, it is not surprising that Ca2+ ion, in conjunction with Mg2+, is a major
component of the dissolved solids in water.
Hardness Classification of Water
Hardness (ppm CaCO3)

Classification

< 15 ppm
15 ppm 50 ppm
50 ppm 100 ppm
100 ppm 200ppm
> 200 ppm

Very soft water

Soft water
Medium hard water
Hard water
Very hard water

The concentration of the hardening ions in a water sample is commonly expressed as

though the hardness is due exclusively to CaCO3. The unit for hardness is mg CaCO3/L,
which is also ppm CaCO3.
In this experiment a titration technique is used to measure the combined Ca2+ and Mg2+
concentration in a water sample. The titrant is EDTA, the disodium salt of
ethylenediaminetetraacetic acid (abbreviated Na2H2Y).
In aqueous solution Na2H2Y dissociates into Na+ and H2Y2 ions. The H2Y2 ion reacts
with the hardening ions, Ca2+ and Mg2+, to form very stable complex ions, especially in a
solution buffered at a pH of about 10. An ammonia-ammonium ion buffer is often used for
this pH adjustment in the analysis. A special indicator called Eriochrome Black T (EBT) is

20

used to detect the endpoint in the titration. EBT forms complex ions with Ca2+ and
Mg2+ions, but binds more strongly to Mg2+ ions. Because only a small amount of EBT is
added, only a small quantity of Mg2+ is complexed; no Ca2+ ion is complexed to EBT,
therefore most of the hardening ions remain free in solution. The EBT indicator is blue
in solution but the [Mg-EBT]2+ complex ion is red.
Mg2+(aq) + EBT(aq)
blue

[Mg-EBT]2+(aq)
red

There even before any H2Y2 titrant is added for the analysis, the solution is red. As H2Y2
titrant is added, it complexes with the free Ca2+ and Mg2+.
Ca2+(aq) + H2Y2 (aq)

CaY2 (aq) + 2H+(aq)

Mg2+(aq) + H2Y2 (aq)

MgY2 (aq) + 2H+(aq)

Once the H2Y2 complexes all of the free Ca2+ and Mg2+ from the water sample, it then
removes the Mg2+ from the [Mg-EBT]2+ complex; the solution turns from red back to blue
colour of the indicator, and the endpoint is reached.
[Mg-EBT]2+(aq) + H2Y2 (aq)
red

MgY2 (aq) + 2H+(aq) + EBT(aq)

blue

For the endpoint to appear Mg2+ must be present; therefore a small amount of MgY2 is
usually added to the buffer solution. The added Mg2+ does not affect the amount of H2Y2
used in the analysis because an equimolar amount of Na2H2Y is also added.
The concentration of chloride ions in water sample can be determined by Mohr
precipitation titration. The water sample is titrated with standard silver nitrate solution and
sodium chromate as indicator. A small amount of calcium carbonate is added to change
the pH of the water sample to basic condition.

APPARATUS
Volumetric flask
Burette with stand
Pipette
Beaker
250 mL Erlenmeyer flasks
REAGENTS
Na2EDTA
Standard Ca2+ solution
Mg/EDTA solution
Ammonia-ammonium ion buffer solution
Eriochrome Black T
0.05M AgNO3 solution
Sodium chromate, Na2CrO4
Calcium carbonate, CaCO3

21

PROCEDURE
I: Determination of the hardness of a water sample
Part IA: To prepare a standard 0.01M Na2EDTA solution
1. Measure about 1.25 g ( 0.01 g) of Na2EDTA (molar mass = 372.24 g/mol); transfer it
to a 250 mL volumetric flask containing about 200 mL of distilled water and stir to
dissolve. Dilute to the mark on the volumetric flask with distilled water.
2. Prepare a burette for titration. Rinse a burette with the Na2EDTA solution and then fill.
Record the initial volume ( 0.02 mL) of the solution.
3. Pipette out 25.0 mL of the standard Ca2+ solution provided into a 250 mL Erlenmeyer
flask, and record its molar concentration. Then add 2 mL of the buffer (pH = 10)
solution, 5 mL of Mg/EDTA solution, and 5 6 drops of EBT indicator. Titrate the
standard Ca2+ solution with the Na2EDTA solution; swirl continuously. Near the
endpoint, slow the rate of addition to drops; the last few drops should be added at 3 5
seconds intervals. The solution changes from red to purple to blue; the solution is blue
at the endpoint.
4. Repeat the titration twice, then calculate the concentration of the Na2EDTA solution.
Part IB: Analysis the hardness of water sample
1. Obtain about 100 mL of a water sample. If the water sample is too turbid, you will
need to gravity filter the sample before the analysis, and if the sample is acidic, add
1M NH3 until it is basic to litmus.
2. Pipette out 25 mL of the water sample into a 250 mL Erlenmeyer flask, add 2 mL of
the buffer (pH = 10) solution, 5 mL of Mg/EDTA solution, and 5 6 drops of EBT
indicator, then titrate with the Na2EDTA solution till the endpoint is reached. Repeat
the titration thrice and then determine the hardness of the water sample.
II: Determination of the concentration of chloride ion
1. Rinse a clean burette with some standard 0.05M AgNO3 solution, and then fill. Record
the initial volume ( 0.02 mL) of the solution.
2. Pipette 25 mL of the water sample into an Erlenmeyer flask. Add 3 5 drops of
sodium chromate solution (yellow colour) and a piece of pea size calcium carbonate.
3. Titrate the water sample slowly with the standard 0.05M AgNO3 solution from the
burette with constant swirling. At the beginning, a white precipitate, AgCl is formed.
As soon as all the Cl ions have been precipitated, a red precipitate, Ag2CrO4, starts
being formed. Stop the titration when the endpoint is reached, that is the red colour
remains.
4. Repeat the titration twice and then determine the concentration of the chloride ions
present in the water sample.
QUESTIONS
1. Why was it necessary to add a small amount of magnesium/EDTA complex to the
calcium sample before titration?
2. A 25 mL sample of well water for chloride determination requires 34.32 mL of
0.05012 M AgNO3 solution to reach a sodium chromate endpoint. Calculate the
concentration of chloride ion in the water sample.
22

Experiment 3: Water Analysis: Hardness Of Water And Chloride

Ion/Chlorine Ion Concentration
Data Sheet
Name:
Student No.:
Date:
_______________________________________________________________________
I: Determination of the hardness of a water sample
Part IA: To prepare a standard 0.01M Na2EDTA solution
Molarity of Ca2+ solution: ________________________________
Particulars

Trial 1

Trial 2

Trial 3

2+

Volume of standard Ca solution used (mL)

Final burette reading (mL)
Initial burette reading (mL)
Volume of Na2EDTA used (mL)
Molarity of Na2EDTA solution (M)
Average molarity of Na2EDTA solution (M)

Part IB: Analysis the hardness of water sample

Particular

Trial 1

Final burette reading (mL)

Initial burette reading (mL)
Volume of Na2EDTA used (mL)
Moles of Na2EDTA used (mol)
Volume of water sample (mL)
Concentration of sample (M)
Mean concentration of sample (M)

23

Trial 2

Trial 3

II: Determination of the concentration of chloride ion/chlorine ion

Particular

Trial 1

Final burette reading (mL)

Initial burette reading (mL)
Volume of AgNO3 used (mL)
Moles of AgNO3 used (mol)
Volume of water sample (mL)
Concentration of sample (M)
Mean concentration of sample (M)

24

Trial 2

Trial 3

EXPERIMENT 4: REDOX TITRATION ASCORBIC ACID

INTRODUCTION
The human body does not synthesize vitamins; therefore the vitamins that we need for
catalyzing specific biochemical reactions are gained only from the food that we eat. We
are generally aware that vitamin C can be obtained from citrus fruits, but it can also be
obtained from a variety of fresh fruits and vegetables. However, storage and processing
causes vegetables to lose a part of their vitamin C content. Cooking leaches the watersoluble vitamin C from the vegetables; in addition, the high temperatures accelerate its
degradation by air oxidation. Therefore to maximize the intake of vitamin C, only fresh
fruits or vegetables should be consumed.
Vitamin C, also called ascorbic acid, is one of the more abundant and easily obtained
vitamins in nature. It is a colourless, water-soluble acid that, in addition to its acidic
properties, is a powerful biochemical reducing agent, meaning it readily undergoes
oxidation, even from the oxygen of the air.
Even though ascorbic acid is an acid, its reducing properties are used in this experiment to
analyze its concentration in various samples. There are many other acids present in foods
that would interfere with an acid analysis and not permit us to selectively determine the
ascorbic acid content. The equation for the oxidation of ascorbic acid is
C6H8O6(aq) + H2O(l)

C6H8O7(aq) + 2H+(aq) + 2e

In analysing for ascorbic acid, the sample is dissolved in water and treated with a
measured excess of iodate ion, IO3 , a strong oxidizing agent; in an acidic solution
containing an excess of iodide ion, I , IO3 converts to I3 (red-brown), a milder oxidizing
agent (Equation 1).
IO3 (aq) + 8I (aq) + 6H+(aq)

3I3 (aq) + 3H2O(l) ------ (Equation 1)

Some of the I3 then oxidizes the ascorbic acid (Equation 2) present in the sample.
I3 (aq) + C6H8O6(aq) + H2O(l)

C6H8O7(aq) + 3I (aq) + 2H+(aq) ---- (Equation 2)

The remaining I3 (the excess, xs) is titrated with a standard thiosulfate, S2O32 , solution,
producing the colourless I and S4O62 ions (Equation 3).
(xs)I3 (aq) + 2S2O32 (aq)

3I (aq) + S4O62 (aq) ------ (Equation 3)

Therefore the difference between the I3 generated initially and that which is titrated in
excess is a measure of the ascorbic acid content of the sample. The stoichiometric point is
detected using starch as an indicator. Just prior to the disappearance of the red-brown I3 in
the titration, starch solution is added; this forms the deep-blue ion, [I3starch] . The
addition of the S2O32 titrant is continued until the [I3starch] has been reduced to I , the
solution appears colourless at the end point.

25

In this experiment you are required to prepare and standardize a Na2S2O3 solution using
solid KIO3 as a primary standard. The standard solution is then used to analyze for
ascorbic acid in the samples provided.

APPARATUS
250-mL volumetric flask
250-mL Erlenmeyer flask
50-mL burette with stand
Glass stirring rod
Beaker
Muslin
Filter funnel

REAGENTS
Potassium iodate, KIO3
Sodium thiosulfate, Na2S2O3
Potassium iodide, KI
Starch solution
0.5 M H2SO4
Sodium hydrogen carbonate, NaHCO3
Vitamin C tablets
Fresh fruit juices

PROCEDURE
Part A: A Primary Standard 0.01 M Potassium Iodate, KIO3, Solution
1. Measure about 0.5 g ( 0.001 g) of KIO3 on weighing paper, transfer the solid to a 250
mL volumetric flask, dissolve and dilute to the mark.
2. Calculate and record the molar concentration of the KIO3 solution.
Part B: A Standard 0.1 M Na2S2O3 Solution
1. Dissolve about 6 g ( 0.001 g) of Na2S2O35H2O with distilled water and dilute to 250
mL. Stir until the salt dissolves.
2. Properly prepare a clean 50 mL burette and fill it with the Na2S2O3 solution, drain the
tip of air bubbles, and read and record the initial volume ( 0.02 mL).
3. Pipette 25 mL of the standard KIO3 solution into a 250 mL Erlenmeyer flask and add
about 1 g ( 0.01 g) of solid KI. Add about 5 mL of 0.5 M H2SO4 and 0.1 g of NaHCO3
(The NaHCO3 reacts in the acidic solution to produce CO2, providing an inert
atmosphere above the solution and minimizing the possibility of the air oxidation of I
ions).
4. Immediately begin titrating with the Na2S2O3 solution. When the red-brown solution
(due to I3 ) changes to a pale yellow colour, add 2 mL of starch solution. Stirring
constantly, continue titrating slowly until the blue colour disappears.

26

5. Repeat the procedure twice by rapidly adding the Na2S2O3 solution until 1 mL before
the endpoint point. Add the starch solution and continue titrating until the solution is
colourless.
6. Calculate and record the molar concentration of the Na2S2O3 solution.
Part C: Sample Preparation
(a) Vitamin C tablet
1. Read the label to determine the approximate mass of vitamin C in each tablet. Measure
( 0.001 g) the fraction of the total mass of a tablet that corresponds to 100 mg of
ascorbic acid.
2. Dissolve it in a 250 mL Erlenmeyer flask with 40 mL of 0.5 M H2SO4 (Remember that
vitamin tablets contain binders and other material that may be insoluble in water do
not heat in an attempt to dissolve it), and then add about 0.5 g NaHCO3.
3. Proceed immediately to Part D.
(b) Fresh fruit sample
1. Filter about 120 mL of freshly squeezed juice through several layers of muslin.
2. Measure the mass ( 0.01 g) of a clean, dry 250 mL Erlenmeyer flask. Add about 100
mL of the filtered juice and again determine the mass.
3. Add 40 mL of 0.5 M H2SO4 and 0.5 g NaHCO3, and then proceed immediately to Part
D.
Part D: Vitamin C analysis
1. Pipette 25.0 mL of the standard KIO3 solution (from Part A) into the sample solution
from Part C and add 1 g of KI.
2. Add about 5mL of 0.5 M H2SO4 and 0.1 g NaHCO3. Titrate the excess I3 in the
sample with the standard Na2S2O3 solution as described in Part B.4. Read and record
the final burette reading ( 0.02 mL).
3. Repeat the analysis twice in order to complete the three trials.
4. Calculate the percent of ascorbic acid in the sample.
QUESTIONS
1. Explain why cooked fruits and vegetables have lower vitamin C content than fresh
fruits and vegetables.
2. If the blue colour does not appear when the starch solution is added during the
titration, should you continue titrating or discard the sample? Explain.
3. 177.42 mL of a fruit juice contains 32% of the recommended daily allowance of
vitamin C (equal to 60 mg). How many mL of the fruit juice will provide 100% of the
recommended daily allowance?

27

Experiment 4: Redox Titration Ascorbic Acid

Data Sheet
Name:
Student No.:
Date:
_______________________________________________________________________

Part A: A Primary Standard 0.01 M Potassium Iodate, KIO3, Solution

Mass of KIO3 (g)
Moles of KIO3 (mol)
Molarity of standard KIO3 solution (M)

Part B: A Standard 0.1 M Na2S2O3 Solution

Particular

Trial 1

Volume of KIO3 solution (mL)

Moles of KIO3 titrated (mol)
Moles of I3 generated (mol)
Final burette reading (mL)
Initial burette reading (mL)
Volume of Na2S2O3 added (mL)
Moles of Na2S2O3 added (mol)
Molarity of Na2S2O3 solution (M)
Average molarity of Na2S2O3 solution (M)

28

Trial 2

Trial 3

Part C & D: Sample Preparation & Vitamin C analysis

Sample name: ________________________________________
Particular

Trial 1

Mass of sample (g)

Volume of KIO3 solution added (mL)
Moles of KIO3 added (mol)
Total moles of I3 generated (mol)
Final burette reading (mL)
Initial burette reading (mL)
Volume of Na2S2O3 added (mL)
Moles of S2O32 added (mol)
Moles of I3 titrated with S2O32 (mol)
Moles of I3 reduced by C6H8O6 (mol)
Moles of C6H8O6 in sample (mol)
Mass of C6H8O6 in sample (g)
Percent of C6H8O6 in sample (%)
Average percent of C6H8O6 in sample (%)

29

Trial 2

Trial 3

EXPERIMENT 5: GRAVIMETRIC DETERMINATION OF

NICKEL(II) ION
INTRODUCTION
The percentage of nickel in a sample may be determined gravimetrically by precipitation
with the organic reagent dimethylglyoxime (DMG). DMG is a bidentate ligand. Nickel is
often added in small amount during the production of steel; the precipitation analysis with
DMG is especially useful in this situation. DMG is a complexing agent, forming a
characteristic bright red coordination compound with nickel ion. In addition to the
gravimetric determination of nickel, DMG is also often used as a spot test to detect the
presence of nickel in a sample on a qualitative basis. DMG does precipitate a few other
metal ions like platinum, but the bright red colour of the Ni(II)/DMG precipitate is distinct
from others.
The precipitate produced by DMG with nickel(II) ion contains two DMG species
complexed per nickel ion and consists of 20.32% nickel by weight. The precipitate is very
fluffy and has a very low density, which makes it somewhat difficult to handle in any great
quantity. For this reason, the practical use of DMG in nickel analyses is restricted to
samples in which the percent of nickel is rather small. As an organic reagent, DMG is not
very soluble in water and is provided as a solution in alcohol. The presence of volatile
alcohol, combined with the nature of the Ni/DMG precipitate itself, causes the precipitate
to creep up the sides of the funnel used for its filtration. Caution must be exercised while
transferring the precipitate to the filter funnel to prevent its loss. Because DMG is a weak
acid, the precipitate of nickel ion is somewhat sensitive to pH. Before precipitation, the
solution is buffered at basic pH with ammonia.

APPARATUS
Sintered glass filtering funnel
Suction filtration apparatus
Oven (110oC)
Filter paper
Hot plate
Beaker 600 mL

REAGENTS
Nickel sample
pH paper
1% dimethylglyoxime solution in alcohol
6 M ammonia solution
Tartaric acid

30

PROCEDURE
Part A: Preparation and Precipitation of the Nickel Sample
1. Clean a 600-mL beaker, rinse with distilled water.
2. Weigh out about 0.5 g of the unknown nickel sample into the beaker. Record the
weight to the nearest mg (0.001 g).
3. Add about 150 mL of distilled water to the beaker.
4. Add approximately 0.2 g of tartaric acid. Stir to dissolve the solids. (The tartaric acid
forms soluble, stable metal complexes with any other metal ions that might be present
in a real sample and will prevent these other metals from precipitating with the nickel
in this determination)
5. Adjust the pH of the sample to between 8 and 9, using 6 M ammonia solution drop by
drop. Do not dip pH paper into the sample. Rather, remove a single drop of the
solution with a stirring rod, and touch the drop to the pH paper.
6. Heat the sample on a hot plate to approximately 80oC. After the sample has been
heated, remove the beaker from the hot plate.
7. Slowly and with constant stirring add 20 mL of 1% dimethylglyoxime solution. An
intensely red, fluffy precipitate should form at this point. If no precipitate forms,
chances are the pH of your solution has not been correctly adjusted.
8. Allow the precipitate to settle until a layer of clear liquid is visible at the top of the
beaker.
9. To ensure that precipitation is complete, add 2 3 drops of 1% DMG to the clear
layer. If no additional red precipitate forms, then you can assume that all of the nickel
has precipitated. If additional precipitate does form at this place, add 2 3 mL
additional 1% DMG and allow the precipitate to settle again. Then test the supernatant
liquid with another single drop of 1% DMG.
10. Allow the sample beaker to cool to room temperature.
Part B: Filtering the Nickel/Dimethylglyoxime Precipitate
As it is being filtered, the Ni/DMG precipitate has a tendency of creeping up the sides of
the filtering funnel. For this reason, never fill the filtering funnel more than half full at
any time.
1. Weigh a piece of filter paper.
2. When the sample beaker have cooled to room temperature, set up filtering funnel into
the suction filtration apparatus. Fit the funnel with the weighed filter paper. Squirt the
filter paper with distilled water to moisten it.
3. Turn on the suction, and slowly begin pouring the supernatant liquid from sample into
the funnel. When most of the liquid has been transferred, gradually begin transferring
the red precipitate. Remember not to fill the funnel more than half full at any time.
4. When the bulk of the precipitate has been transferred from the beaker to the funnel,
use small portions of distilled water to transfer any remaining particles from the
beaker.
5. When all the precipitate has been transferred to the funnel, increase suction to the
maximum level, and draw air through the precipitate for two minutes to help dry it.

31

6. Transfer the filter paper with the precipitate to a watch glass and dry the filter paper
with precipitate in the oven for at least one hour to remove all moisture. Make sure that
the oven temperature is near 100oC.
7. When the precipitates have dried completely, weigh the filter paper with the precipitate
(to the nearest milligram).
8. Compare your data with two other groups in your laboratory.
9. Calculate the mean % Ni in the original sample.
Part C: Calculations
The Ni/DMG precipitate is known to contain 20.32% nickel by weight.
The mass of nickel present in a given amount of precipitate is then:
(mass of precipitate)

(0.2032)

The percent nickel in the original sample is then:

Mass of nickel
Mass of sample taken

100%

QUESTIONS
1. The percentage of nickel in the Ni/DMG precipitate is 20.32%. Using the atomic mass
of Ni and the formula mass of DMG, derive the percentage.
2. During the experiment, it was necessary to test the nickel sample for complete
precipitation of nickel by adding a single drop of DMG to the supernatant solution.
What error would be introduced into the percent Ni determined for a sample if not all
the nickel had been precipitated?
3. The DMG id provided as a solution in alcohol. Why is alcohol, rather than water, used
as the solvent?

32

Experiment 5: Gravimetric Determination of Nickel(II) Ion

Data Sheet
Name:
Student No.:
Date:
_______________________________________________________________________

Particulars

Sample 1

Mass of original sample (g)

Mass of filter paper (g)
Mass of filter paper with
precipitate (g)
Mass of precipitate (g)
Mass of Ni in precipitate (g)
% Ni in original sample
Mean % Ni in original sample

33

Sample 2

Sample 3

EXPERIMENT 6: THIN-LAYER CHROMATOGRAPHY

INTRODUCTION
The word chromatography means colour-writing. The name was chosen when the method
was first used to separate coloured components from plant leaves. Chromatography in its
various forms is perhaps the most important known method of chemical analysis of
mixtures.
Thin-layer chromatography (TLC) is a simple technique that can be used to separate
mixtures into the individual components of the mixture. It uses a thin coating of
aluminium oxide (alumina) or silica gel on a glass slide or plaster sheet to which the
mixture to be resolved is applied. A single spot of the unknown mixture to be analyzed is
applied about 1 cm from the end of a strip of a TLC slide. The slide is then placed in a
shallow layer of solvent or solvent mixture in a jar or beaker. Since the coating of the TLC
slide is permeable to liquids, the solvent begins rising by capillary action.
As the solvent rises to the level at which the spot of mixture was applied, various effects
can occur, depending on the constituents of the spot. Those components of the spot that
are completely soluble in the solvent will be swept along with the solvent front as it
continues to rise. Those components that are not at all soluble in the solvent will be left
behind at the original location of the spot. Most components of the unknown spot mixture
will take an intermediate approach as the solvent front passes. Components in the spot that
are somewhat soluble in the solvent will be swept along by the solvent front, but to
different extents, reflecting their specific solubilities. By this means, the original spot of
mixture is spread out into a series of spots or band, with each spot representing one single
component of the original mixture.
The separation of a mixture by chromatography is not solely a function of the solubility of
the components in the solvent used. The TLC slide coating used in chromatography is not
inert, but consists of molecules that may interact with the molecules of the components of
the mixture being separated. Each component of the mixture is likely to have a different
extent of interaction with the slide coating. This differing extent of interaction between the
components of a mixture and the molecules of the support forms an equally important
basis for the separation. The TLC slide coating adsorbs molecules on its surface to
differing extents, depending on the structure and properties of the molecules involved.
To place a TLC separation on a quantitative basis, a mathematical function called the
retention factor, Rf, is defined:
Rf

distance traveled by spot

distance traveled by solvent

The retention factor depends on what solvent is used for the separation and on the specific
composition of the slide coating used for a particular analysis. Because the retention
factors for particular components of a mixture may vary if an analysis is repeated under
different conditions, a known sample is generally analyzed at the same time as an
unknown mixture on the same slide. If the unknown mixture produces spots having the Rf
values as spots from the known sample, then an identification of the unknown components
has been achieved.

34

Indicators are organic compounds that are typically used to signal a change in pH in
acid/base titration analyses. Such indicators are dyes that exist in different coloured forms
at different pHs, and change in colour of the indicator is the signal that the titration
analysis is complete.
In this experiment, you will perform a thin-layer chromatographic analysis of a mixture of
the dyes bromocresol green, methyl red, and xylenol orange. These dyes have been chosen
because they have significantly different retention factors, and a nearly complete
separation should be possible in the appropriate solvent system. You will also investigate
the effect of the solvent on TLC analyses, by attempting the separation in several different
solvent systems.

APPARATUS
TLC slides
Beakers
Parafilm
Micropipette
Ruler
Pencil

REAGENTS
Methyl red
Xylenol orange
Bromocresol green
Acetone
Ethyl acetate
Hexane
Ethanol

PROCEDURE
1.
2.

Clean and dry six beakers to be used as the chambers for the chromatography.
Prepare mixtures of the solvents below, in the proportions indicated by volume, and
transfer each to a separate beaker. Cover the beakers with parafilm after adding the
solvent mixture, and label the beakers with the identity of the mixture it contains.
Acetone : hexane (3 : 2)
Ethyl acetate : hexane (3 : 2)
Acetone : ethyl acetate (1 : 1)
Acetone : ethanol (1 : 1)
Ethyl acetate : ethanol (1 : 1)
Hexane : ethanol (1 : 1)

3.

Prepare six TLC slides by marking lightly with pencil (not ink) a line across both the
top and bottom of the slide. Do not mark the line too deeply or you will remove the
coating of the slide. See Figure 6.1

35

Figure 6.1
4.

5.

6.

7.

8.
9.

On one of the lines on each slide, mark four small pencil dots (to represent where the
spots are to be applied). Above the other line on each slide, mark the following letters:
R (methyl red), X (xylenol orange), G (bromocresol green) and M (mixture). See
Figure 6.1.
Obtain small samples of the ethanol solutions of the three dyes and the mixture.
Apply a single small droplet of the appropriate dye and the mixture to its pencil spot
on each of the TLC slides. Use a separate micropipette for each dye and the mixture.
Keep the spots of dye and the mixture as small as possible.
Allow the spots on the TLC slides to dry. Then gently lower one of the TLC slides,
spots downward, into one of the solvent systems. Be careful not to wet the spots, or to
slosh the solvent in the beaker. Do not move or disturb the beaker after adding the
TLC slide. Carefully cover the beaker with parafilm.
Allow the solvent to rise on the TLC slide until it reaches the upper pencil line. Then
remove the TLC slide and quickly mark the exact solvent front before it evaporates.
Mark the TLC slide with the identity of the solvent system used for development. Set
the TLC slide aside to dry completely.
Repeat the process using the additional TLC slides and solvent systems. Be certain to
mark each slide with the solvent system used.
Determine Rf for each dye in each solvent system and record. Keep the TLC slides
and staple to the lab report for this experiment.

QUESTIONS
1.

Why is it important to keep the spots applied to TLC slide for chromatography as
small as possible?

2.

Why is it necessary to keep the beaker used for chromatography tightly covered with
parafilm while the solvent is rising through the TLC slide?

3.

Of the solvents used, some were very polar (eg. acetone, ethanol) while others were
very nonpolar (eg. hexane). Does the polarity of the solvent of the various solvent
mixtures seem to affect the completeness of the separation of dyes? Why might this
be so?

36

Experiment 6: Thin-layer Chromatography

Data Sheet
Name:
Student No.:
Date:
_______________________________________________________________________
For each of the solvent mixtures studied, calculate Rf for each of the spot:
Acetone : hexane

Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

Ethyl acetate : hexane Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

Acetone : ethyl acetate Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

37

Acetone : ethanol

Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

Ethyl acetate : ethanol Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

Hexane : ethanol

Distance travelled by solvent front _______________________

Distance travelled by spot

Calculated Rf

Methyl red

_____________________

_______________________

Xylenol orange

_____________________

_______________________

Bromocresol green

_____________________

_______________________

Which solvent mixture gave the most complete resolution of the three dyes? Which
solvent mixture gave the poorest resolution?
________________________________________________________________________

38