NORTHERN CARIBBEAN UNIVERSITY

DEPARTMENT of BIOLOGY, CHEMISTRY and

ALLIED HEALTH SCIENCES

LABORATORY MANUAL

CHEM202: ANALYTICAL CHEMISTRY II

Prepared by: Dr Nicole White

TABLE OF CONTENTS

PAGE
 • TABLE OF CONTENTS 1
• CHEMISTRY LABORATORY REGULATIONS AND SAFETY

PRECAUTIONS 2

• EXPERIMENT 1: THE DETERMINATION OF ACID CONTENT

IN VINEGARS 8
• EXPERIMENT 2: THIN LAYER CHROMATOGRAPHY 9
• EXPERIMENT 3: PREPARATION OF A STANDARD SILVER
NITRATE SOLUTION 13
• EXPERIMENT 4: DETERMINATION OF CHLORIDE CONTENT
BY THE MOHR METHOD 14
• EXPERIMENT 5: CHLORIDE CONTENT IN DIFFERENT
SOURCES OF WATER 16
• EXPERIMENT 6: GRAVIMETRIC DETERMINATION OF CHLORIDE
IN A SOLUBLE SAMPLE 18
• EXPERIMENT 7: DETERMINATION OF CALCIUM BY
DISPLACEMENT TITRATION 20
• EXPERIEMENT 8: DETERMINING COPPER CONTENT IN SOIL

USING ATOMIC ABSORPTION SPECTROSCOPY 22

CHEMISTRY LABORATORY REGULATIONS AND SAFETY PRECAUTIONS

Reference: http://www.sciencebyjones.com/safety_rules.htm

Thanks to the Flinn Scientific Safety Rules for much of the above.

Compiled by Nicole White (PhD) 2008

General Guidelines

1. Conduct yourself in a responsible manner at all times in the laboratory.

2. Be familiar with your lab assignment before you come to lab. Follow all written and
verbal instructions carefully. If you do not understand a direction or part of a procedure,
ask the teacher before proceeding.

3. Never work alone. No student may work in the laboratory without an instructor present.

4. When first entering a science room, do not touch any equipment, chemicals, or other
materials in the laboratory area until you are instructed to do so.

5. Do not eat food, drink beverages, or chew gum in the laboratory. Do not use laboratory
glassware as containers for food or beverages.

6. Perform only those experiments authorized by the instructor. Never do anything in the
laboratory that is not called for in the laboratory procedures or by your instructor.
Carefully follow all instructions, both written and oral. Unauthorized experiments are
prohibited.

7. Safety goggles and aprons must be worn whenever you work in lab. Gloves should be
worn whenever you use chemicals that cause skin irritations or need to handle hot
equipment. Wear older clothes that cover the maximum amount of skin.

8. Observe good housekeeping practices. Work areas should be kept clean and tidy at all
times. Bring only your laboratory instructions, worksheets, and/or reports to the work
area. Other materials (books, purses, backpacks, etc.) should be stored in the classroom
area.

9. Know the locations and operating procedures of all safety equipment including the first
aid kit, eyewash station, safety shower, spill kit, fire extinguisher, and fire blanket. Know
where the fire alarm and the exits are located.

10. Be alert and proceed with caution at all times in the laboratory. Notify the instructor
immediately of any unsafe conditions you observe.

11. Dispose of all chemical waste properly. Never mix chemicals in sink drains. Sinks are to
be used only for water and those solutions designated by the instructor. Solid chemicals,
metals, matches, filter paper, and all other insoluble materials are to be disposed of in the
proper waste containers, not in the sink. Check the label of all waste containers twice
before adding your chemical waste to the container. Cracked or broken glass should be
placed in the special container for “Broken Glass.”

12. Labels and equipment instructions must be read carefully before use. Set up and use the
prescribed apparatus as directed in the laboratory instructions provided by your teacher.
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13. Keep hands away from your face, eyes, mouth, and body while using chemicals. Wash
your hands with soap and water after performing all experiments. Clean (with detergent
powder), rinse, and dry all work surfaces and equipment at the end of the experiment.

14. Experiments must be personally monitored at all times. You will be assigned a laboratory
station at which to work. Do not wander around the room, distract other students, or
interfere with the laboratory experiments of others.

15. Students are never permitted in the science storage rooms or preparation areas unless
given specific permission by their instructor.

16. Know what to do if there is a fire drill during a laboratory period; containers must be
closed, gas valves turned off, fume hoods turned off, and any electrical equipment turned
off.

17. If you spill acid or any other corrosive chemical on you skin or clothes immediately wash
area with large amounts of water (remember that small amounts of water may be worse
that no water at all). After this get the teacher’s attention. The spill kit will be used for
spills on floor or counter-top.

18. At the end of the laboratory session see that: a) main gas outlet valve is shut off b) the
water is turned off c) desk top, floor area, and sink are clean d) all equipment is cool,
clean, and arranged.

Clothing

19. Any time chemicals, heat, or glassware are used, students will wear laboratory goggles.
There will be no exceptions to this rule! Contact lenses should not be worn in the
laboratory unless you have permission from your instructor.

20. Dress properly during a laboratory activity. Long hair, dangling jewelry, and loose or
baggy clothing are a hazard in the laboratory. Long hair must be tied back and dangling
jewelry and loose or baggy clothing must be secured. Shoes must completely cover the
foot. No sandals are allowed.

Accidents and Injuries

21. Report any accident (spill, breakage, etc.) or injury (cut, burn, etc.) to the instructor
immediately, no matter how trivial it may appear.

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22. If you or your lab partner is hurt, immediately yell to get the instructor's attention.
Everyone should turn off burners and prepare to help if needed.

23. If a chemical should splash in your eye(s), immediately flush with running water from the
eyewash station for at least 20 minutes. Notify the instructor immediately.

Handling Chemicals

24. All chemicals in the laboratory are to be considered dangerous. Do not touch, taste, or
smell any chemical unless specifically instructed to do so. The proper technique for
smelling chemical fumes (when instructed to do so by the teacher) is to gently fan the air
above the chemical toward your face. Breathe normally.

25. Check the label on chemical bottles twice before removing any of the contents. Take only
as much chemical as you need. Smaller amounts often work better than larger amounts.
Label all containers and massing papers holding dry chemicals.

26. Never return unused chemicals to their original containers.

27. Never use mouth suction to fill a pipette. Use a pipette bulb or pipette filler.

28. Acids must be handled with extreme care. ALWAYS ADD ACID SLOWLY TO
WATER, with slow stirring and swirling, being careful of the heat produced, particularly
with sulfuric acid.

29. Handle flammable hazardous liquids over a pan to contain spills. Never dispense
flammable liquids anywhere near an open flame or source of heat.

30. Never take chemicals or other materials from the laboratory area.

31. Take great care when transferring acids and other chemicals from one part of the
laboratory to another. Hold them securely and in the method demonstrated by the teacher
as you walk.

Handling Glassware and Equipment

32. Inserting and removing glass tubing from rubber stoppers can be dangerous. Always
lubricate glassware (tubing, thistle tubes, thermometers, etc.) before attempting to insert it
in a stopper. Always protect your hands with towels or cotton gloves when inserting glass
tubing into, or removing it from, a rubber stopper. If a piece of glassware becomes
"frozen" in a stopper, take it to your instructor for removal.

33. When removing an electrical plug from its socket, grasp the plug, not the electrical cord.
Hands must be completely dry before touching an electrical switch, plug, or outlet.

Compiled by Nicole White (PhD) 2008

34. Examine glassware before each use. Never use chipped or cracked glassware. Never use
dirty glassware. Do not immerse hot glassware in cold water; it may shatter.

35. Report damaged electrical equipment immediately. Look for things such as frayed cords,
exposed wires, and loose connections. Do not use damaged electrical equipment.

36. If you do not understand how to use a piece of equipment, ask the instructor for help.

Heating Substances

37. SHOULD THE BUNSEN BURNER GO OUT, IMMEDIATELY TURN OFF THE
GAS AT THE GAS OUTLET VALVE. If you wish to turn off the burner, do so by
turning off the gas at the gas outlet valve first, then close the needle valve and barrel.
Never reach over an exposed flame. Light gas burners only as instructed by the teacher.

38. Never leave a lit burner unattended. Never leave anything that is being heated or is visibly
reacting unattended. Always turn the burner or hot plate off when not in use.

39. You will be instructed in the proper method of heating and boiling liquids in test tubes.
Do not point the open end of a test tube being heated at yourself or anyone else.

40. Heated metals, glass, and ceramics remain very hot for a long time. They should be set
aside to cool on a wire gauze and then picked up with caution. Use tongs or heat-
protective gloves if necessary. Determine if an object is hot by bringing the back of your
hand close to it prior to grasping it.

GENERAL NOTES
• The Abstract should provide a less-than-150-word summary of the entire work: the

Compiled by Nicole White (PhD) 2008

 purpose, procedure, key results, and their significance should all be briefly addressed in
this essential part of your report. The Abstract is not the place to introduce the experiment
or describe the underlying principles in any detail. Stated in another way, the paper really
begins with the Introduction, not the Abstract. Most scientists write the Abstract after they
have written the rest of the paper, since it summarizes the work described. Never present
material in the Abstract that you have not also presented somewhere in the main body of
the report.

• The Introduction should describe the specific goals of your experiment. What have you
analyzed, and why? Briefly discuss some of the key ideas. You are not required to use
other references, but you are welcome to.

• The Procedure should provide a concise description of how the experiment was actually
conducted. Note important observations (especially events that likely introduced error)
and highlight any deviations from the instructions in the handout you do not need to
include drawings of any apparatus used in the experiment unless you feel it will aid your
discussion.

Calculating acid concentrations from reagent stock

The molarity (cm) and percentage (cp) relationship depends on the density of solution (d) along
with the molecular mass (MM) of the dissolved substance. Two equations depicting the
interconversion of these two are as follows:

cm = cp × d / (100% × MM)

Reference:
− http://www.trimen.pl/witek/calculators/stezenia.html
− http://www.macalester.edu/~kuwata/classes/2005-
06/Chem%20222/Pb%20in%20Soil%20Lab%202006.pdf

Compiled by Nicole White (PhD) 2008

 GENERAL LAB EQUIPMENT

Reference
1. http://images.google.com/imgres?imgurl=http://physics.gallaudet.edu/classes/chemappa.gif&imgrefurl=https://sites.google.com/a/ggwo.org/ggca_science_lab/Home/chemistry/chemistry-equipment--
safety&h=851&w=680&sz=140&hl=en&start=2&um=1&usg=__UvvQ2Dnd800KdbfMqNavZKP_T_Y=&tbnid=KsuthEI8w4YH9M:&tbnh=145&tbnw=116&prev=/images%3Fq%3Dlab%2Bequipme
nt:%2Bchemistry%26um%3D1%26hl%3Den%26rls%3Dcom.microsoft:en-gb:IE-SearchBox%26rlz%3D1I7RNWM%26sa%3DN
2.http://images.google.com/imgres?imgurl=http://visual.merriam-webster.com/images/science/chemistry/laboratory-equipment_2.jpg&imgrefurl=http://visual.merriam-
webster.com/science/chemistry/laboratoryequipment_2.php&h=384&w=550&sz=55&hl=en&start=3&um=1&usg=__3DXNsK64lkexY4xcVrs89SaC0PU=&tbnid=VenjDu5mM2Rq5M:&tbnh=93&tbn
w=133&prev=/images%3Fq%3Dlab%2Bequipment:%2Bchemistry%26um%3D1%26hl%3Den%26rls%3Dcom.microsoft:en-gb:IE-SearchBox%26rlz%3D1I7RNWM%26sa%3DN
3. http://visual.merriam-webster.com/images/science/chemistry/laboratory-equipment_1.jpg 4. http://bioweb.wku.edu/courses/Biol121/Carbo/pipets.png
5. http://www.naugraexport.com/glass/front.jpg 6. http://www.science-laboratory-equipment.com/product-images/1059.jpg
7. http://www.unisciencelab.com/unisciencelab/laboratory/images/tongs_1.jpg 8. http://z.about.com/d/chemistry/1/0/D/U/potferri.jpg

Compiled by Nicole White (PhD) 2008

 EXPERIMENT 1

THE DETERMINATION OF ACID CONTENT IN VINEGARS

Aim
− To determine the acid contents in various brands of vinegars.

Introduction
Vinegar or French for sour wine is formed by aerobic bacteria oxidizing grain alcohol to acetic
acid and water. More generally, vinegar can be defined as a solution composed of acetic acid
(HC2H3O2), water, and, perhaps, other substances. To be sold in stores as vinegar, this solution
must contain at least four grams of acetic acid per 100 ml of solution. For the titration of the
vinegar in this experiment the following specific reaction will be used to calculate the acetic acid
content of the vinegar sample:
H3C2O2H(aq) + NaOH(aq) → H2O(l) + C2H3O2Na (aq).

% w/v = (mass of solute/volume of solution)×100%.

Procedure
Keep flask closed when not in use as the acidity of bottled vinegar tends to decrease on exposure
to air. Record the brand of vinegar used.
1. Standardize the ~ 0.1NaOH solution using potassium hydrogen phthalate.
2. Pipette 25 mL of the unknown into a 250 mL volumetric flask and dilute to the mark with
distilled water.
3. Pipette 50 mL aliquot of vinegar into a 250 mL conical flask. Add 50 mL of water.
4. Titrate against the NaOH solution.
Titrations are to be done in triplicate.

Treatment of data
• Determine the concentration of NaOH used.
• Determine the w/v% vinegar in each sample.

Questions
1. Does the vinegar studied meet the commercial law specification of a minimum of 4 g of
acetic acid/100 ml of vinegar?
2. Has the vinegar supplier truthfully reported the percent acidity? Why is it a good idea to
rinse the burette with the NaOH solution, instead of with water, before filling it at the
start of the titration?

Reference
− http://www.baruch.cuny.edu/wsas/departments/natural_science/chemistry/chm_1000/vinegar.pdf

Compiled by Nicole White (PhD) 2008

 − Skoog, D.A.; west, D.M.; Holler, F.J. Foundations of analytical Chemistry, 7th Ed.;
Saunders College Publishing: 1991
EXPERIEMENT 2

THIN LAYER CHROMATOGRAPHY

Aim
− To investigate the separation of mixtures with thin-layer-chromatography and calculate R f
values
− To determine the composition of various over-the-counter analgesics

Introduction
Since the investigation of chemically pure materials has provided the basis for understanding
chemistry, it is critical that chemists be able to separate mixtures of substances as they are often
found in nature. Thin layer chromatography is a special application of adsorption in which a thin
layer of adsorbent supported on a flat surface is utilized instead of a column of adsorbent. The
most commonly used adsorbent is silica gel and the flat surface is a plain glass plate (a cut piece
of sheet glass). All forms of chromatography depend upon the distribution (or separation) of
solute particles between a moving phase (a gas or liquid; you'll be using a liquid moving phase in
this procedure) and the stationary phase (a liquid or solid; in this procedure you will be using a
solid stationary phase.
The separation of the components of a mixture depends on adsorption-desorption equilibria
between compounds adsorbed on the solid stationary phase and in the moving liquid phase. The
extent of adsorption of a single component depends upon the polarity of the molecule, the
activity of the adsorbent, and the polarity of the mobile liquid phase. The actual separation of the
components in a mixture is dependent on the relative values of the adsorption-desorption
equilibrium constants for each of the components in the mixture. In general, the more polar a
functional group in the compound is, the more strongly it will be adsorbed on the surface of the
solid phase. If the components are colored, they can be located readily, but more often they are
invisible and must be located by other means. Illumination with ultra-violet light will excite
fluorescence in many compounds.

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Because each chromatogram will be developed under slightly different conditions (slightly more
or less solvent, slightly different temperatures, differing size and concentration of the origin line
applications), the general practice in TLC is to place known and unknown compounds on the
same chromatogram. The unknowns can then be easily identified by comparison with the known
samples that have been run under the exact same conditions on the same chromatogram. A few
characteristics that remain consistent from run to run can be used to compare different
chromatograms. The generally accepted method for making such comparisons is to compute the
Rf value (meaning "Relationship to the Front") of the spots. This is easily calculated by:

Rf = Distance moved by compound

Distance moved by solvent system

(Note that a more polar compound will more strongly adsorb to the polar stationary phase
and will have a lower Rf value)

The means, therefore, available to identify the components of a TLC mixture are:
• Migration identical to that of a standard or reference (known) compound
• Intrinsic fluorescence like that of a reference compound
• Staining behavior like that of a reference compound (Staining characteristics may be
concentration-related. Therefore, a spot of lower concentration may have a lighter color than a
spot of higher concentration)
• The Rf value that is identical to that of a reference compound. This Rf value may be obtained
from direct measurement or obtained from literature.

You will be using TLC to separate and identify the components of some familiar mixtures of
organic compounds – over-the-counter analgesic drugs. Several of these compounds have related
structures, but are different enough so that they can be separated.
Aspirin is acetylsalicylic acid – made from the esterification reaction of salicylic acid with acetic
anhydride. It was first used in ancient times, when the Greek physician Hippocrates had his
patients chew willow bark to ease pain and reduce fever. It was later discovered that willow bark
contains salicin, a derivative of the molecule we know as aspirin today. Aspirin is an analgesic
(pain reliever) as well as an antipyretic (fever reducer). Its action in both cases, analgesic and
antipyretic, is due to the ability of aspirin to inhibit prostaglandins, which are chemicals in the
body involved in sending pain messages to the brain. Any analgesic doesn't really stop the ache
or pain; an analgesic just keeps the brain from hearing about the pain. Aspirin also can inhibit the
clotting of blood, so sometimes it is prescribed for lowering the risk of heart attack and stroke
(cerebral vascular accidents) by lowering the formation of blood clots that can dislodge and plug
arteries or veins in the heart and brain.
Acetaminophen is structurally related to aspirin, and it is an analgesic and an antipyretic, but
does not reduce inflammation like aspirin does. It also does not promote gastrointestinal upset
like aspirin does, so it can be taken in larger doses over longer periods of time.
Caffeine is often found in preparations of over-the-counter analgesics, but there is little evidence
that caffeine enhances the effects of aspirin. Caffeine is a mild stimulant.
Salicylamide is also structurally related to the aspirin, and is sometimes used in combination
with aspirin.

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Ibuprofen is also an analgesic (but not used in this procedure). It only bears a slight structural
resemblance to aspirin, but may be superior to aspirin in the reduction of inflammation. It can
also reduce fever and relieves mild pain, but does not appear to be better than aspirin for these
effects.
Procedure
Use the ethyl acetate + 0.5% acetic acid solvent for this section. Be careful; don't
shine the UV light in your eyes.

1. Obtain on thin layer chromatography plate from your instructor. Handle the plate by the
edges only. With a pencil and ruler, lightly mark a line about 1 cm from the narrow edge
of the plate (not the wide edge of the plate).
2. Divide the plate into six equal portions. Six solutions will be spotted on each plate
(keeping the spots as small as possible, approximately 1-2 mm in diameter) – a solution
of each of four standards/reference solutions (acetaminophen, aspirin, caffeine,
salicylamide) will be applied to the plate, as well as a combined solution of REV 01/06
11 all four standards/references together, so it can be determined how the compounds run
when together in solution. You will also spot your assigned unknown on the plate. Apply
each of the six solutions once; then spot your unknown a second time, allowing the spot
to dry before placing the other spot on top of it. It might be easiest to apply the solutions
in alphabetical order, to prevent confusion. Apply each substance twice to plate on the
same spot.
3. Dry the plates in an 80 °C oven for 2-3 minutes (especially if the weather is humid). The
chromatograms will be developed in a mixture of ethyl acetate/acetic acid.
4. While the plates are drying in the oven, prepare the chromatography chamber. You will
use your 400-mL beaker as a development chamber. Place enough of the solvent
(ethylacetate/acetic acid) into the bottom of the chromatography so that the depth of
solvent is enough to cover the bottom of the chamber, but not enough to touch the applied
spots.
5. Insert the spotted, dried plate into the prepared chamber and cover the chamber with a
plastic sandwich bag. Do not move the jar during development. Watch the progress of the
solvent front on the plate. When the solvent front is about 0.5-1.0 cm from the top of the
plate, remove the plate from the chamber and QUICKLY mark the solvent front level
with a pencil line across the top of the chromatogram before the solvent evaporates. After
marking the solvent front, allow the chromatogram to dry. Place the used solvent in the
proper waste container. Clean the chamber (a 400-mL beaker) with soap and water. Rinse
the beaker with deionized water as a final rinse.
6. Visualize the plate under short-wave UV light, and mark any spots (either fluorescent or
dark spots) that are seen with a pencil on your chromatogram. Then place the
chromatogram into a jar with a few iodine crystals, cover the jar and let stand until
brownish-yellow spots develop. Remove the plates and outline the spots with a pencil.
7. Notice the placement of the compounds:
Analgesic Rf Value
Acetaminophen (A) 0.476
Salicylamide (S) 0.738
0.362
Aspirin (Asp) 0.619
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 Ibuprofen (I) 0.785
Caffeine (C) 0.362

Aspirin and salicylamide run at about the same Rf value; the way they are told apart is by the
visualization process. If both compounds are present, both visualization steps will show the
presence of the proper compounds.

Treatment of data
• Make a drawing of this chromatogram in your laboratory notebook, indicating the proper
colour and shape of each spot.
• Measure the distance from the origin line to the solvent front. Measure all distances in
centimeters and measure to the nearest 0.01 cm (two decimal places). Then measure the
distance from the origin line to the center of each spot (again, to the nearest 0.01 cm).
Write these distances on the drawing in your notebook.
• Using these measured distances, calculate the Rf of these spots.
• Determine the components

Questions
1. What would happen if the solvent layer in your development chamber is too deep, so that
the origin lines of your chromatograms are submerged in it?
2. Using an appropriate source, draw the structural formula of the following compounds in
the spaces provided above each name. a) aspirin b) ibuprofen c) acetaminophen
d) caffeine

Reference
- http://www.ipfw.edu/chem/112/kimble/2-Thin%20Layer%20Chromatography.pdf
- http://delloyd.50megs.com/labscripts/TLC.html
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 - http://bama.ua.edu/~kshaughn/ch338/handouts/TLC-exp1.pdf

EXPERIMENT 3

PREPARATION OF A STANDARD SILVER NITRATE SOLUTION

Aim
− To utilize proper analytical techniques in preparation of a standard solution

Introduction
Most precipitation titrations make use of a silver nitrate solution as titrant. It offers several other
advantages as well. It is non-hygroscopic, in contrast to silver fluoroborate and silver
perchlorate. It is relatively stable to light. Finally it dissolves in numerous solvents. The nitrate
can be easily replaced by other ligands, rendering AgNO3 versatile. Treatment with solutions of
halide ions gives a precipitate of AgX (X = Cl, Br, I).

CAUTION
As with all silver salts, silver nitrate is toxic and corrosive. Brief exposure to the chemical will
not produce immediate or even any side effects other than the purple skin stains, but with more
exposure, side effects will become more noticeable. It is also very poisonous and can cause
burns. Long-term exposure may cause eye damage. Short contact can lead to deposition of black
silver stains on the skin. Besides being very destructive of mucous membranes, it is a skin and
eye irritant.

Procedure
1. Weigh approximately 16.9 g of AgNO3 and dry at 110 oC for no more than an hour.
Prolonged heating causes partial decomposition of AgNO3. Cool to room temperature in a
dessicator.
2. Weigh the bottle and contents to the nearest 0.0001 g.
3. Transfer the bulk to a 1000 mL volumetric flask using a funnel.
4. Cap the weighing bottle and re-weigh.
5. Rinse the funnel thoroughly.
6. Dissolve the AgNO3. Fill flask to the mark.

Treatment of data
• Calculate the molar concentration of this solution.

Reference
− http://en.wikipedia.org/wiki/Silver_nitrate#Preparation
− Skoog, D.A.; west, D.M.; Holler, F.J. Foundations of analytical Chemistry, 7th Ed.;
Saunders College Publishing: 1991
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 SAVE SOLUTION FOR NEXT 3 LABS. STORE IN A DARK PLACE.

EXPERIMENT 4

DETERMINATION OF CHLORIDE CONTENT BY THE MOHR METHOD

Aim
− To find the percentage of Cl- ions in an unknown compound.

Introduction
Based on the chemical reaction, the chloride ions precipitated as silver chloride in the final
titration experiment.
AgNO3 + NaCl → AgCl + NaNO3
The Mohr method uses the indicator potassium chromate. This method determines the chloride
ion concentration of a solution by titration with silver nitrate. As the silver nitrate solution is
slowly added, a precipitate of silver chloride forms.
Ag+(aq) + Cl–(aq) → AgCl(s)
The end point of the titration occurs when all the chloride ions are precipitated. Then additional
chloride ions react with the chromate ions of the indicator, potassium chromate, to form a red-
brown precipitate of silver chromate.
2 Ag+(aq) + CrO42–(aq) → Ag2CrO4(s)
The Mohr titration should be carried out under conditions of pH 6.5 – 9. At higher pH silver ions
may be removed by precipitation with hydroxide ions, and at low pH chromate ions may be
removed by an acid-base reaction to form hydrogen chromate ions or dichromate ions, affecting
the accuracy of the end point.

Procedure
Dry reagent grade NaCl and unknown in oven for ca. 1 hr. at 110 oC. Cool in a dessicator.
1. Prepare 0.1 M AgNO3 solution. Express the concentration as weight molarity (mm
AgNO3/g of solution)
2. Dissolve 1.0 g K2CrO4 in approximately 20 mL of distilled water.
To standardize AgNO3:
3. Weigh 0.25 g portions of NaCl into 250 mL Erlenmeyer flask. Dissolve in 100 mL
distilled water.
4. Add small quantities of NaHCO3 until effervence stops.
5. Add ~2 mL K2CrO4 solution to NaCl solution.
6. Titrate against AgNO3 until you observe the first permanent red Ag2CrO4.
7. Determine an indicator blank by suspending a small amount of chloride free CaCO 3 in
100 mL of distilled water containing 2 mL of K2CrO4. Titrate as above.
Unknown sample:
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 8. Collect unknown from instructor.
9. Weigh accurately 0.2 g of sample. Titrate as above.

Treatment of data
• Correct reagent weights for the blank.
• Calculate %Cl- in NaCl.
• Determine % Cl- in unknown.
• Compare your result with the other groups.

Reference
− http://www.neiu.edu/~rdpurtuc/iSchool/chem/quant%20213/Quant-Lab-02.pdf
− Skoog, D.A.; west, D.M.; Holler, F.J. Foundations of analytical Chemistry, 7th Ed.;
Saunders College Publishing: 1991
− http://www.mcm.edu/~simpsong/Quant2.htm
− http://www.chemteach.ac.nz/investigations/documents/chloride_mohr.pdf

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 EXPERIMENT 5

CHLORIDE CONTENT IN DIFFERENT SOURCES OF WATER

Aim
− To determine the percentage of chlorine present in water from various sources

Introduction
Tests for the presence of chloride ion are frequently required when testing water supplies for
chlorination. In the pulp and paper industry, bleaching plants must do extensive analysis for
chlorides before returning waste water to a river or stream. Treatment for chloride removal is
costly and requires some level of technical expertise. In general, water having the highest
quality is reserved for the potable water supply. It meets all international standards for drinking
water and contains a chloride concentration less than 250 mg/L. Industrial water use varies
widely (boiler-feed makeup, process water, truck washing, toilet flushing, etc) and therefore the
water quality also varies widely. Agriculture can generally accept water having lower quality
standards, but the salinity of irrigation water depends upon crop tolerance and rainfall. Irrigation
water having more elevated levels of salinity can be tolerated by most crops in areas having high
rainfall, evenly distributed throughout the year

The procedure for testing for chloride ions in solution is fairly simple. Most chloride tests
involve titration with silver nitrate solution in the presence of chromate ion which acts as an
indicator. Chloride ions can be precipitated with silver nitrate to form insoluble silver chloride.

Ag+(aq)) + Cl- (aq)) à AgCl (s)

Silver chloride is less soluble than silver chromate. Therefore when the chloride has been
removed from the solution the orange-red silver chromate begins to precipitate signaling the end
point.

2 Ag+(aq)) + CrO42- à Ag2CrO4 (s)

In this experiment you will be determining the concentration of Chloride ion in seawater. The
usual concentration is about 1.93% but the value varies slightly depending on the depth and the
location where the sample was collected.

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Procedure
Silver nitrate will stain the skin and can be harmful if taken internally. Potassium
Chromate may be a suspected carcinogen if inhaled or airborne. Avoid all contact with the
skin
1. Weigh the pipette containing the salt water solution. Then put 20 drops of the salt water
solution into a well of the 24 well plate or a small test tube. The well or test tube should
be one quarter to one third full.
2. Carefully add 2-3 drops of the potassium chromate solution and stir with a toothpick.
3. Record the initial mass of the silver nitrate pipette.
4. Add the silver nitrate a drop at a time while stirring with the toothpick or swirling the
solution between each additions.
5. Continue adding silver nitrate a drop at a time until a red precipitate persists throughout
the liquid and cannot be stirred away.
6. For sea water about 20-30 drops will be required. Reweigh the silver nitrate pipette to
determine the mass of AgNO3 added.
7. Repeat experiment for: pool water, tap water, and bottled water.

Treatment of data
• Tabulate you results as follows.
Mass/g Sample data
Initial mass salt water dropper
Salt water dropper after adding water to test
tube
Mass of salt water
Initial mass of the dropper with silver nitrate
Mass of dropper after adding silver nitrate to
test tube
Mass of silver nitrate solution used

• Calculate the mass of salt water used.

• Calculate the mass of silver nitrate used.
• Find the number of moles of silver nitrate used. (Multiply the mass in calculation 2 by
the density of the solution and the concentration of silver nitrate solution in moles per
dm3)
• 5. Find the mass of Chloride ion present
• 6. Find the percentage of Chloride ion in solution

Reference
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 − www.sidsnet.org/docshare/other/20031105152734_CUBA.CASE_STUDY2.doc

− http://lincoln.pps.k12.or.us/pages/lscheffler/ChlorideAnalysis.htm

PREPARE CRUCIBLES FOR NEXT WEEKS EXPERIMENT.

EXPERIMENT 6

GRAVIMETRIC DETERMINATION OF CHLORIDE IN A SOLUBLE SAMPLE

Aim
− To accurately determine the mass of a substance by employing crucibles

Introduction
Gravimetric analysis is a quantitative (i.e. how much?) method of classical analysis. The element
to be determined is isolated in a solid compound of known identity and definite composition. The
chloride content of a soluble salt can be determined by precipitation as silver chloride:

Ag+ + Cl- →AgCl(s)

In most cases, the analyte must first be converted to a solid by precipitation with an appropriate
reagent. The precipitate can then be collected by filtration, washed, dried to remove traces of
moisture from the solution, and weighed. The amount of analyte in the original sample can then
be calculated from the mass of the precipitate and its chemical composition.

Procedure
Preparation of crucibles and preliminary lab work
1. Clean 2 medium porosity sintered glass by allowing them to stand in approximately 5 mL
of concentrated nitric acid for 5 minutes.
2. Using vacuum filtration, draw the acid the crucible.
3. Rinse thoroughly with tap water and finally with distilled water.
4. Allow each crucible and dry to constant weight in an oven at 110 oC. Three constant
readings between 0.2 -0.3 mg are sufficient. Do not touch crucibles with hand once the
drying process has begun.
5. Place crucibles in a dessicator until required for use. Allow it to cool to room
temperature.
6. Transfer unknown to a 400 mL beaker.
7. Dissolve in 100 mL distilled water. Add 2-3 mL of HNO3.
8. Slowly, and with good stirring, add 0.2 mL AgNO 3 to each cold sample solutions until
AgCl begins to coagulate. Use separate stirring rods for each beaker.
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 9. Add an adiitional 3 mL of AgNO3. Heat to almost boiling.
10. Digest for 10 minutes. Add a few drops of AgNO3 to confirm precipitation is complete.
11. If incomplete (more ppt. forms), then add 3 mL of AgNO3, digest, and again test for
precipitation.
12. Cover each beaker and store in a dark place until the next laboratory session

Experiment
13. Decant the supernatant liquids through weighed filtering crucibles (previously prepared).
Wash the precipitates several times with a solution of 2-5 mL of 6 M HNO3 per litre of
distilled water.
14. Filter, using vacuum filtration, liquids through crucibles. Use policemen to dislodge any
particles that adhere to the walls of the beakers. Note which beaker was paired to which
crucible.
15. Dry precipitates in an oven at 110 oC for at least 1 hr. Store the crucibles in a dessicator
while they cool.
16. Determine the mass of crucibles and their contents.
17. Repeat the cycle of heating, cooling, and weighing (30 minute periods) until constant
weights are obtained.
18. Place solids into paper and finally placed in waste provided. All solutions should also be
placed in waste containers.
19. Clean crucibles by repeating steps 1-3.

Treatment of data
• Tabulate all masses, properly records must be kept.
• Calculate the percentage of Cl- in the sample.

Reference
− http://www.earlham.edu/~chem/chem111f03/labs/Experiment%206.pdf
− http://en.wikipedia.org/wiki/Gravimetry
− Skoog, D.A.; west, D.M.; Holler, F.J. Foundations of analytical Chemistry, 7th Ed.;
Saunders College Publishing: 1991

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 EXPERIMENT 7

THE DETERMINATION OF CALCIUM BY DISPLACEMENT TITRATION

Aim
− To determine the metal content in a complex

Introduction
A solution of the magnesium/EDTA complex is useful for the titration of cations that form more
stable complexes than the magnesium complex but for which no indicator is available.
Ethylenediaminetetraacetic acid (EDTA) is a reagent that forms EDTA-metal complexes with
many metal ions (but not with alkali metal ions such as Na+ and K+). The concentration of the
magnesium solution is not important; all that is necessary is that the molar ratio between Mg 2+
and EDTA in the reagent be exactly unity. In alkaline conditions (pH >9) it forms stable
complexes with the alkaline earth metal ions Ca2+ and Mg2+. The EDTA reagent can be used to
measure the total quantity of dissolved Ca2+ or Mg2+ ions in a water sample.

Procedure
Preparation of Solutions
1. Buffer solution, pH 10: Dilute 75 mL of concentrated NH 4OH and 10.5 g NH4Cl- in
sufficient distilled water to give 150 mL of solution.
2. Eriochrome Black T indicator: Dissolve 250 mg of the solid in a solution containing 15
mL of triethaloamine and 5 mL absolute ethanol.
3. Prepare 6M and 0.1 M NaOH solutions in 250 mL volumetric flasks.
4. Prepare 6 M HCl solution in a 100 mL volumetric flask.
5. MgSO4/ EDTA solution: Dissolve 3.72 g EDTA in 50 mL of distilled water and add 2.46
g MgSO4.7H2O. Add a few drops of phenolphthalein indicator and titrate against 0.1 M
NaOH until the white solution turns pink.
6. Blank solution: Mix 10.00 mL distilled water + 2 drops of Erichrome Black indicator + 2
drops of pH 10 buffer solution + 1.00 mL of MgEDTA solution, and titrate dropwise
against 0.01M EDTA.

Standardization of 0.01M EDTA against Ca2+ solution of known concentration

7. Weigh 0.9320g EDTA and mix with 250 mL of distilled water to give 0.01M EDTA.
8. Weigh 0.2503g CaCO3 and mixed it with 5.00 mL 6M HCl + 50.00 mL H2O.
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 9. Heat solution to boil for 3-4 minutes; let solution cool to room temperature.
10. Titrate solution, carefully, against 6 M NaOH.
11. Dilute solution to 250 mL with water
12. Pipette 10mL of new solution and mix it with 2 drops of Erichrome Black indicator + 2
drops of pH 10 buffer solution + 1.00 mL of MgEDTA solution.
13. Titrate the 10.00 mL sample solution against stock 0.01M EDTA.
14. Pipette 20.00 mL of CaCO3 for triplicate titrations against EDTA

Determination of unknown Calcium by displacement titration

15. Weigh 0.5g (0.4999g) of unknown sample and mix with 5.00 mL 6M HCl + 50.00 mL
water.
16. Heat solution to boil for 3-4 minutes; let solution cool to room temperature.
17. Titrate solution against 6 M NaOH.
18. Dilute solution to 250 mL with H2O.
19. Pipette10.00 mL of new solution and mix it with 2 drops of Erichrome Black indicator+ 2
drops of pH 10 buffer solution + 1.00 mL of MgEDTA solution.
20. Titrate, in triplicate, the 10.00 mL sample solution with stock standardized EDTA
solution

Treatment of data
• Calculate EDTA molarity.
• Calculate mass percentage of Ca2+ in unknown sample.

Reference
− http://www.neiu.edu/~rdpurtuc/iSchool/chem/quant%20213/Quant-Lab-05.pdf
− Skoog, D.A.; west, D.M.; Holler, F.J. Foundations of analytical Chemistry, 7th Ed.;
Saunders College Publishing: 1991

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 EXPERIMENT 8

DETERMINING COPPER CONTENT IN SOIL USING ATOMIC ABSORBTION

SPECTROSCOPY

Aim
− To determine the copper concentration in soil

− To understand and apply a calibration curve

Introduction
Atomic absorption spectroscopy (AA or AAS) is a technique for determining the concentration
of a particular metal element in a sample. It is one of the commonest instrumental methods for
analyzing for metals and some metalloids. The technique makes use of absorption spectrometry
to assess the concentration of an analyte in a sample and relies therefore heavily on Beer-
Lambert Law. The analyte concentration is determined from the amount of absorption.
In their elemental form, metals will absorb ultraviolet light when they are excited by heat. Each
metal has a characteristic wavelength that will be absorbed. The AAS instrument looks for a
particular metal by focusing a beam of uv light at a specific wavelength through a flame and into
a detector. The sample of interest is aspirated into the flame. If that metal is present in the
sample, it will absorb some of the light, thus reducing its intensity. The instrument measures the
change in intensity. A computer data system converts the change in intensity into an absorbance.
As concentration goes up, absorbance goes up. The researcher can construct a calibration curve
by running standards of various concentrations on the AAS and observing the absorbances.

Sample treatment
1. Approximately, 5 g of each sample was placed in a 50 mL volumetric flask and the
container filled to the mark with distilled water.
2. The samples were then run using an atomic absorption spectrometer at a wavelength of
324.75 nm. The samples were run in triplicates.

Results
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Absorbance readings for standard copper samples
[Cu]/ mg/L Mean Absorbance (A)
0.000 0.0185
0.502 0.0400
0.993 0.0600
2.005 0.0950
3.999 0.1550

Absorbance readings for unknown copper samples

Sample A
1 0.0434
2 0.0470
3 0.0498
4 0.0491
5 0.0481
6 0.0472
7 0.0653
8 0.0617
9 0.0523

Treatment of data
• Determine the concentration of each of the unknown samples from the calibration curve.
The absorbances have not been corrected for the blank. Make the necessary corrections.

• What are the concentrations of the original samples?

Questions
1. Why were the measurements carried out in triplicates?
2. What is the purpose of the blank run?
3. If you were a field analyst asked to determine the average copper content in a particular
parish, outline the procedure you would use to carry out this investigation.

Reference
− http://en.wikipedia.org/wiki/Atomic_absorption_spectroscopy
− http://elchem.kaist.ac.kr/vt/chem-ed/spec/atomic/aa.htm
− http://www.shsu.edu/~chemistry/primers/AAS.html
− http://www.gmu.edu/departments/SRIF/tutorial/aas/aas.htm

Special thanks to Dr M. Wilson and S.I.R.I.

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