Professional Documents
Culture Documents
The trick with both table manners and lab technique is to learn one method, develop
it into a habit, and refuse to do things any other way, so that the habit becomes
entrenched. In this manual each operation is spelled out in detail the first time you
meet it. The methods are chosen to be "goof-proof" and to work with almost all
substances, almost all of the time. Learn them, and let them become your laboratory
table manners.
You can also avoid trouble by understanding what you are doing and why. If you
have had difficulty in previous labs, be especially careful about prelab preparation.
Your roommate may be able to come into lab unprepared and get away with it, but
do not try it. The extra preparation time you put in will be saved by just one
afternoon your roommate wasted because of a bad day," and you will be ahead.
Last, do not try to beat the clock. People work at different speeds. Since teaching
assistants are paid for the class period, asking them to stay on is an unreasonable
imposition, and legal requirements do not let you work unsupervised. But
sometimes you can come back to another session another afternoon and continue.
Be sure to observe whatever the local tradition is regarding admission to another
section.
At bottom, the idea that scientists have to have exceptional dexterity is a myth. In
practice, experiments are designed to be done with all constraints-dexterity being
one of the more fundamental ones.
When writing a book like this, there is a choice between making it completely
humorless, or allowing some of the personality of the author to come through. I
have chosen the latter to avoid making the text sound like instructions to Form 1040.
I realize that humor is a subtle thing that does not always come across well in
writing, but I very much want you to read and enjoy this manual. Good luck in all
of your scientific endeavors!
Miles Pickering
5. With a disposable dropper, remove most of the liquid, discarding the liquid
into the "mercury waste" container in the hood. Add 2 to 3 mL of distilled water
and bring to the boil. Again, cool, let the solid settle, and discard the waste
solution into the mercury waste container.
6. Add a few drops of distilled water. Use the tip of a disposable dropper to stir
up the precipitate and suck up the solid. Transfer it to a clean 50 mL beaker.
Allow the beaker to stand uncovered for a week in your desk. This will allow
the excess H2O to evaporate.
The following is an example of how the procedure appeared in a student's notebook.
Receive test tube with 30 mg of Hg2+ (poisonous).
Add 1 mL of 0.5 M KI sol. shake 30 seconds.
Solid should form, redissolve. If not completely redissolved, add more KI drop-wise until
solution is transparent.
Add 1 mL of 0.1M Ag+ or 0.1 M Cu2+, agitate.
Heat 5 to 10 min in H2O bath.
Remove liquid with disposable dropper, discard into "Hg waste."
Add 2 to 4 mL of H2O. Heat to boil in H2O bath.
Discard H2O into "Hg waste.
Add few drops H2O, stir up solid with dropper, suck up drop into beaker, let stand one
week.
Observe and record colors.
The previous is a good notebook summary because
1. it is broken up into individual steps and has a check-list quality;
2. all key landmarks are present (for example, "solid should form, redissolve") so
that the student can check the results as the work goes on;
3. hazards are included;
4. there is a reminder to record crucial observations.
This summary could be written on the left-hand half of the page, leaving the other half
page for observations. Alternatively, the student could entitle it To Be Done and then
record measurements and observations at the end under the heading What Was
Done.
Keeping Records
Nowhere is habit so important as in keeping accurate records. A number of otherwise
inexplicable student results are caused by scrambled records, and studies of laboratory
exams have shown that this is a major cause of student failure. Therefore, most
teachers are strict about notebooks this is an attempt to save students from
themselves. Your instructor will almost certainly add to the directions in this manual.
Your notebook must be of an approved type and data must be recorded in ink. It must
be current and consecutive, and all your work must be in one place. In many schools,
3
notebooks are checked on a weekly basis by the teaching assistant, and a persistently
unsatisfactory notebook counts against your grade.
Never, never tear out pages (except carbons): not if you make a mistake, not if you spill
something, never!
Record three classes of data.
1. All of the numbers pertaining to the experiment. Identify all numbers and follow by a
unit. Be sure to record concentrations of stock solutions. This information must be
recorded directly in your notebook, not on scraps of paper. Even if it is transferred
once, there is a danger of, for example, digits being interchanged, so be sure to take
your notebook whenever you go to the balance. Some teachers enforce this by
picking up all scrap paper. Beware!
2. All the observations you have made. This includes smells, colors, and color changes.
It includes the size and shape of solid crystals, abrupt changes in temperature, or
any other phenomena. When in doubt, record it.
3. Any changes in procedure. If you diluted a reagent down from a concentrated stock
solution, record it. If there was a dead fly in your flask when you did a titration,
record it. When in doubt, record it. In many colleges you will be tested on the facts
you have observed. You may be permitted to have your notebook for reference
during the test, so it will be to your advantage if it is complete and readable.
In some colleges, at the end of each period you must turn in a carbon copy of your
notebook for that period. This is not only to keep you honest; it helps you if your
notebook is lost or stolen, because it serves as a record.
WORKING SAFELY
We have done our best to build safety into the design of the experiments and work
areas, but working in the lab is still not as safe as lying in bed. Your instructor will
undoubtedly bring local safety rules to your attention. The ones listed below, which
are universal, serve as a starling point.
1. Wear eye protection at all times. If you ponder for a moment what would
happen to the rest of your college and professional career if you were accidentally
blinded this very period, I am sure you will agree that the discomfort of safety
glasses is a small price to pay for some added insurance. In any case, you must
wear eye protection because it is mandated by federal regulations (29 CPR
1910.133). If you refuse to wear eye protection, you cannot come to lab. It is that
simple.
2. Do not wear contact lenses to lab. If you wear contact lenses, and chemicals get
into your eyes, much more severe damage will result.
3. Wear shoes that cover your feet (not open-toed shoes or sandals). This, too, is
mandated by federal regulation. Do not wear shorts or skirts in the lab. Uou
should wear long pants. Shoes and clothes provide a first line of defense against
spills. Old blue jeans are a good choice for lab clothes. They resist chemicals, and
stains on them are not noticeable. If all else fails, they can be thrown out at the end
of the semester. You will also be required to wear a lab apron when working in the
lab. You are required to wear a lab apron when working in the lab.
4. Do not eat, or drink, in the lab. As the alchemist Paracelsus remarked, "All things
are poisonous, for there is nothing without poisonous, qualities. It is only the dose
which makes a thing poison." These words are as true now as in the sixteenth
century. If you need to eat, or drink, do it in the hall. Although few chemicals are
absorbed through your skin, avoid touching chemicals whenever possible. Wash
your hands at the end of the lab period.
5. Be aware that burns and fires are serious hazards. Some of the substances that
you work with burn very easily. Also note that hot glassware looks very much the
same as cold glassware, but its effect on your skin is very different!
6. Do not perform any unauthorized experiments. The difference between using
potassium chlorate and chloride may be an explosion!
7. Report even trivial injuries to your teaching assistant or lab supervisor. What
seems minor can have major complications.
8. Work in the lab only if you are being supervised. You can work only during your
scheduled period, unless other arrangements are made, and even then you may
work only in the presence of a teaching assistant.
9. Know the location of emergency exits, emergency showers, eye-wash fountains,
and fire extinguishers. Always know two different ways out of a lab. Do not put
book bags or other things on the floor, where they might impede escape. In a real
emergency, follow the instructions of teaching assistants or supervisors. Move, do
not argue!
10. Do not, as a rule, bring visitors into most laboratories. Ask permission before
bringing them, and see that they obey the rules. You can always talk in the hall.
11. Do not listen to music through earphones. (You want to be able to hear
instructions in an emergency.)
If you are, or might be, pregnant, consult your instructor about postponing your lab.
All chemicals known to cause birth defects at this writing have been excluded, but so
little is known about this topic that a conservative approach seems appropriate. The
threat of birth defects from toxic agents is most serious in the first 3 months; it seems
that the hazard is much less later.
When you notice a fellow student who is not following roles, remind him or her.
Almost certainly, your fellow student would rather be reminded by you than by a
teaching assistant or faculty member! If you are the one being reminded, thank the
person doing it and be glad you did not find out "by accident."
Using Proper Lab Etiquette
Lab work can be made faster, pleasanter, and simpler for everybody if your class can
use peer pressure to encourage people to observe some rules that fall into the category
of etiquette. For example, do not remove tools and reagents that are to be shared. Do
not stick droppers or other tools into stock solution bottles. If you spill something,
clean it up. At the end of the period, sponge off your bench, so that the next user does
not have to start by cleaning up a puddle of colorless liquid, wondering, "Is it water or
sulfuric acid?"
USING THE TOOLS OF THE TRADE
Bunsen burners, beakers and test tubes are a few of the tools you will use for the
following experiments.
Running a Reaction
To run a reaction you will need to mix the chemicals (called reagents) and possibly to
beat them. These basic techniques are covered in this section.
How to Adjust a Bunsen Burner
In order to accurately perform an experiment using a Bunsen burner, the flame needs
the proper mixture of gas and air. The flame will be noisy or blow out if there is too
much air or not enough gas. A smoky or yellow flame indicates that there is too much
gas or not enough air. Regulate the air by adjusting the barrel. Control the gas with a
needle valve in the burner base.
How to Set Up a Beaker or Flask for Boiling
Place a piece of wire gauze on a tripod, then place the beaker or flask on top of the wire
gauze. The gauze helps support the beaker and also spreads the heat. To stop the heat,
remove the lighted burner from under the flask, then let the contents of the container
stop boiling before attempting to pour or remove. Use beaker tongs or crucible tongs to
grasp a hot object.
Bumping
Boiling may be uneven and explosive. This phenomenon, known as bumping, is
controlled by adding boiling chips. These act as nuclei, giving the vapor bubbles a
place to form so that the liquid does not superheat. Before adding boiling chips, always
let the hot liquid cool below the boiling point. Otherwise a burst of boiling will occur
when the chips hit the liquid, often scattering the liquid out of the container.
How to Work With a Test Tube
Many of the experiments in this lab manual are done in test tubes. To mix solutions, or
to get a solid to dissolve, do not seal the test tube with your thumb and invert it. This
will contaminate the solution and may be dangerous to tour skin. Rather, grasp the top
of the test tube firmly and then tap rapidly with a finger from the other hand. Use a
circular stroke, letting your finger quickly slide down the outside of the tube (Fig. 1). If
you do this correctly you can create a vortex in the test tube that will mix it efficiently.
Figure 1 Mixing solutions in a Test Tube.
Never heat a test tube directly. Always immerse it in a hot water bath. Otherwise the
liquid may bump and fly out of the test tube.
Measuring Weight
Many of the measurements in this manual depend on the analytical balance, which is
potentially one of the most accurate instruments available to you. Learn to use this
balance well. Your teaching assistant will explain how to use the particular model of
balance available in the labs
How to Use the Analytical Balance
Whenever you are asked to weigh out a solid, you will be given a range of weights.
Any weight in this range will work, as long as you know exactly how much you
actually weigh out.
Weigh by difference. Put the solid in a vial, put the top on (to keep out moisture and
dust), and take a reading. Then transfer a small amount of the solid into a beaker or
flask and reseal the vial. Pour the solid directly into the beaker or flask; do not use a
spatula lest some of the solid stick to it. Reweigh the sealed vial. If the amount
transferred is not sufficient, transfer more to the beaker and reweigh the vial. If you
have added too much, clean out the beaker and start over. Do not transfer the solid
back to the weighing vial lest you contaminate the entire sample.
Remember that the analytical balance is a very precise instrument. Always record all of
the available decimal places. You can round off the number later.
For precise work, do not weigh hot objects on the balance. A hot object creates
convection currents of air that cause the reading to appear light. Although many books
suggest holding objects only with tissue, this is unnecessary unless you have very
sweaty fingers. Hugging the sample in the palm of your hand is not recommended and
will lead to a steady downward drift as the moisture evaporates.
The area in and around the balance must be kept clean. Spills are expected, and you
will never be penalized if you clean them up. However, leaving a mess for the next
student is discourteous and dangerous.
How to Use the Beam Balance
The beam balance is used for rough measurements only and should not be used unless
specified explicitly in this book. Directions vary from balance to balance, and your
instructor will advise you in using yours.
MEASURING VOLUME
Learning to do good volume measurements is essential, and not difficult.
How to Clean Glassware
Soap and water will remove almost all of the stains you will have on glassware (except
as noted for KMnO4) in this course, although it may take some scrubbing. Labels can
be removed by soaking in hot water and then scraping with a spatula, using the same
motion as that for peeling an apple.
It is not necessary to use a great excess of soap. Excessive amounts are difficult to
remove and require many rinsings, which takes time. Do the final rinsing with a small
amount of distilled water (just enough to thoroughly wet the inside, but not enough to
fill or even halfway fill the container).
How to Take Solutions from Stock Bottles
Use a graduated cylinder to take solutions from stock bottles. Pour the stock solution
from the stock bottle into the graduated cylinder, until you have taken slightly more
than you need. Take the cylinder to your desk, and remove the excess with a dropper.
Never, never pour the excess back into the stock bottle. Never, never put any object
(for example, a dropper) into the bottle. If you encounter difficulty with a bottle, get
assistance from your TA.
Remember this as the "one-way" rule: Things come out of stock bottles but never go
into them!
Do not take stock bottles to your desk. Also, if bottles have been placed in the hoods,
leave them there. The hood helps confine poisonous, smelly, or corrosive spills.
How to Use Volumetric Flasks
A volumetric flask must never be heated, because it will probably break. Even if it does
not break, the volume calibration will be destroyed. Dissolve solids before adding them
to the flask, rinse container used for the solutions with a wash bottle, and add the
rinsings to the flask. Shake thoroughly, then fill up to the mark (a circular ring etched
on the neck) and shake again. You cannot mix too much, but each year many students
have difficulties because they did not mix enough! If you fill the flask past the mark,
discard the solution and start over (Fig. 2).
Do not store solutions in volumetric flasks. The solution may cause the stopper to
"freeze, that is, to become solidly wedged in the flask. If this happens, get assistance
for frozen stoppers.
TC
20 C
Pyrex USA No. 5640
The inscription above appears on a typical volumetric flask. What does it mean? The
top line, A, means "Class A according to the National Bureau of Standards. The next
line means "To Contain at 20 C, 250 mL with an absolute uncertainty of 0.12 mL."
The third line means "made of Pyrex glass (a registered trademark for a type of
borosilicate glass) made in the United States, Catalog No. 5640." Note that the old
abbreviation ml is now being replaced by mL.
ACCURATELY MEASURING VOLUME WITH A BURET
Beads of water form on the inside of an unclean buret when it is wet, as a result of an
invisible layer of grease. This may interfere with the accuracy of the measurement,
because drops remaining on the side of the buret will be counted as if they had been
delivered. Soapy water and a buret brush are sufficient for cleaning the buret.
After rinsing the buret, including the top, several times with water, rinse it with a little
of the solution you will use. This prevents accidentally diluting the solution. Fill the
buret over the sink, which you should be able to do without a funnel and without
spilling. (Funnels can introduce contamination, unless they, too, are with a little of the
solution to be transferred.) Run some of the liquid until the tip of the buret is filled and
the meniscus is on scale. It is a waste of time to start at 0.00 mL.
10
To read the buret, look at the bottom of the liquid meniscus and then guestimate the
last place, so that you have a reading with two places after the decimal. The volume
delivered is the difference between the first reading and the final reading, and is
measured in milliliters (mL). The numbers increase reading down the buret, rather than
up as on a thermometer (Fig. 3).
Figure 3 Buret Reading
If you cannot remove all the air from the tip, you may get a false reading when it fills
up later. To remove an air bubble, hold the buret over the sink, open the valve, and
shake once vertically. Ask the teaching assistant for help if you are having trouble
PIPETTING
Two sorts of pipettes are available: the measuring pipette, which is often used in
clinical medicine and has graduations along the side, and the transfer pipette, which
has a bulge in the center. The latter is much more accurate. To use a transfer pipette,
follow this procedure.
1. Clean it thoroughly to prevent beads of water from clinging to the inside.
These beads count as delivered, even though they remain inside.
2. Hold the pipet with one hand and the pipet bulb with the other, out the air
from the bulb, then loosely attach it to the top of the pipet until the liquid has
been pulled up above the mark. Quickly remove the bulb and put your finger
over the top of the pipette.
3. Slowly let the level fall until the bottom of the meniscus just touches the line.
Release the top of the pipette, and let it deliver the solution into the vessel.
Touch the tip to the side of the container, and hold it there for a few seconds
after it looks as if all the contents have drained out. Do not blowout the
pipette or put your mouth on it at any time.
11
12
13
CAUTION:
Potassium Hydroxide, KOH, and Sulfuric Acid, H2SO4, are corrosive and must not be
allowed to touch the skin, eyes or clothes. Methanol, CH3OH, and Ethanol C2H5OH
burns easily; keep fire and flames away when working with them.
Preparation of Kalinite:
1. Accurately weigh between 0.20 - 0.25 g of alloy chips and transfer to medium size
test tube. Add 10 mL of 1.4 M KOH solution (be careful not to get any KOH solution
on your skin. If you do, immediately wash with water), and warm the solution in a
warm water bath. Be careful not to BOIL this solution. After 20 -30 min remove the test
tube from the water bath and allow the test tube to cool. Remember to record all
observations (i.e. color changes, evolution of gases).
2. Gravity filter the cooled solution into a small beaker. To the filtrate carefully add 5
mL of 9 M H2SO4 (be careful not to get any H2SO4 solution on your skin. If you do,
immediately wash with water). If a precipitate still remains after all the acid has been
added, warm the solution again, until all the solid is dissolved.
3. Get 10 mL of methanol from the hood. Before bring the methanol back to your
bench, make sure there are no flames in the immediate vicinity. Methanol Burns! Place
the beaker in an ice bath, add the 10 mL of methanol to the solution, and allow the
solution to cool for 10-20 min. During this time crystals will form. Collect the crystals
by vacuum filtration using a Buchner funnel. Your TA will demonstrate how to
perform a vacuum filtration. You can obtain a Buchner funnel from the stockroom.
Purification of the Kalinite:
The crude kalinite contains a small excess of K2SO4 impurities, and these crystals will be
purified via recrystallization from water. Recrystallization is the most common method
for purifying a solid material. This method take advantage of the differences in
solubility of a compound in hot and cold solvent. The impure solid is dissolved in the
minimal amount of hot solvent and as the solutions cools the pure product crystallizes
from solution leaving the impurities behind in solution.
Heat some distilled water. Place your Kalinite in a small 50 mL beaker, and add the
hot water drop wise with a disposable pipet until all the solid dissolves. It may be
necessary to warm the solution of water and Kalinite if it becomes too cool. When all
the solid is in solution, remove from the heat and allow the solution too cool. You can
place the beaker in an ice bath to speed the cooling process, but if you allow the
solution to cool slowly you will get larger crystals. Collect the solid on a Buchner
funnel. Obtain an exact weight of the dry crystals.
14
% yield =
actual yield
100
theorectical yield
The theoretical yield is the number of grams of product that would be obtained if the
reaction went to 100% completion and produced only the desired product. The
theoretical yield forthis reaction can be determined as follows:
a. Since the aluminum is the limiting reagent, calculate the number of moles of
aluminum that you started with. The alloy chips used in this experiment are 92
% pure aluminum.
b. Since the mole ratio of Al : Kalinite is 1:1, calculate the number of grams of
Kalinite produced if the reaction went to 100% completion. Molecular weight
for Kalinite is 474.39 g/mol
" wt Al chips %" 1 mole KAl(SO 4 ) 2 12 H 2O %" 474.39 g %
theoretical yield = 0.92$
'$
'$
'
# atomic wt Al &#
&# mole &
1 mole Al
From the above formula and your data, calculate the percent yield for this reaction?
3. The ion Fe3+ is very similar in size to Al3+ and can replace it in the crystal lattice of
kalinite, so that kalinite with an iron impurity is common. How can one produce pure
kalinite from contaminated kalinite? (Hint: dissolve the kalinite in water then add
something that will precipitate the iron.)
15
COOH
COONa
+ NaOH
+ H2O
COOK
COOK
NaOH solution can be calculated from the recorded volume of NaOH solution required
to neutralize a know weight of KHP. This method of determining precise concentration
of an unknown NaOH solution is called Standardization.
The procedure requires on knowing exactly when the neutralization has occurred. This
problem is solved by using an indicator, a dye that will undergo a rapid, reversible
molecular rearrangement in solution. There is a conspicuous color difference between
the molecular forms in acidic and in basic solutions, and the colors are so bright that
only a little of the indicator need be used. A single drop of NaOH solution will convert
some of the dye from its acidic form to its basic form so the first excess of NaOH not
used to neutralize the acid will convert the dye to its basic form, which is brightly
colored. This result indicates that neutralization reaction has occurred. This point is
known as the end point of the titration.
16
Last, be sure, when filling, to write down the concentration and any other information
appearing on the stock bottle label.
17
CAUTION:
Sodium Hydroxide, NaOH, is corrosive and must not be allowed to touch the skin,
eyes or clothes. Clean up any spills, and rinse skin with water. Inform your TA of
any spills.
Part A: How to prepare the NaOH solution.
A stock solution of concentrated NaOH will be provided. Add about 30 mL of this
solution to 900 mL of distilled water in a clean plastic bottle, stopper and mix well.
Mixing well means shaking the bottle for at least 1 full minute with, as much vigor as
you can muster more shaking does no harm. The diluted NaOH, while poisonous, is
not nearly as corrosive as the concentrated solution.
Part B: Standardizing the NaOH solution with KHP.
Take the plastic vial containing your KHP to the balance with a conical flask. Weigh
the vial to get the exact weight. Then transfer between 0.5000 g and 0.8000 g of solid to
the first flask, recording the final weight of the vial, to obtain the exact amount
transferred by difference. Add about 50 to 100 mL of deionized water to the conical
flask. Warm to dissolve, and add between 3-5 drops of phenolphthalein indicator.
Fill a buret with NaOH, check for air bubbles, and then record the initial reading. Now
add NaOH solution from the buret to the flask, quickly at first. The pink color of the
indicator will persist for an increasingly longer time, and will finally spread through
the solution. It is useful to swirl with your dominant hand and control the volume on
the buret with your nondominant hand. Continue adding the solution drop-wise as
you approach the end point.
You will probably need to repeat this procedure several times before you find two
satisfactory end points, but it goes quickly after the first run. Compute the exact NaOH
concentration, and label your bottle. Keep the NaOH in your desk. You will need it for
subsequent work.
Calculating the Concentration of Sodium Hydroxide
The calculation depends on the important fact that one mole of KHP will neutralize
exactly one mole of NaOH. At the endpoint then, the moles NaOH will equal the moles
of KHP.
1. The molecular weight of KHP is 204.22 g/mol. Compute the number of moles of
KHP weighed out.
The volume must be expressed as liters (1 mL is 0.001 L) if the units are to work out.
The concentration of NaOH is simply the number of moles of KHP per the number of
liters of NaOH solution run out of the buret.
18
The concentration or Molarity (M) of the NaOH solution can be calculated, and the unit
for Molarity is mol/Liter:
Molarity NaOH =
%
weight KHP "
1
$
'
204.22 g/mol # vol. NaOH &
2. Do the uncertainty calculations. Start by calculating the upper and lower limits for
your best run. If uncertain as to which is best, choose the last one because practice may
cause an improvement.
For KHP samples weighed by difference, the uncertainties in
weight of KHP will be 0.0010 g. The two worst cases for the Molarity of the NaOH
solution will be
%
upper - limit weight KHP "
1
$
'
204.22 g/mol
# lower - limit vol. NaOH &
%
lower - limit weight KHP "
1
$
'
204.22 g/mol
# upper - limit vol. NaOH &
3. Are the two concentrations identical within uncertainty? If not, shake the NaOH,
refill the buret, and do a third run to see if you can obtain agreement. (The shaking is
recommended only because incomplete mixing is the most common student error.)
4. To determine if two titrations experiments agree within uncertainty, look at the range
of concentrations. One titration of NaOH should fit inside the range of uncertainty just
calculated. For example if your range is 0.0876M to 0.0888M and your first value is
0.0879M, you have agreement. If not, at most one run can be correct.
Part C: A Proof of Method test of your NaOH Solution
The purpose of this little experiment is (1) to provide you with a yardstick by which to
measure your titration technique before you embark on the titration of real unknowns,
and (2) to give you an idea of what the lab practical exam will be like.
You will be given a sample of an unknown acid and are to report the apparent
molecular weight. Remember to record the Unknown Acid Number written on the vial
label in your notebook.
Weigh a sample of 0.5000 g to 1.5000 g of unknown acid into a conical flask, recording
the exact weight. Add 50 100 mL de-ionized H2O to the conical flask, and swirl to
dissolve the solid. Add between 3-5 drops of phenolphthalein indicator.
You handle the unknown acid in the same way you handled the KHP. The first run
will probably be overshot, but weigh out another sample. Initially you can go quickly
until you're close, then drop-wise.
19
There is no uncertainty analysis required for the Unknown Acid titration. Your TA will
tell you where to check your value of the Unknown Acid molecular weight. If your
agrees, then you have accurately determined the molecular weight. If your value
value
does not agree, then perform another titration.
For the Report
1. Make a table containing the net weights of KHP and net volume of NaOH solution
used in each titration. You should include data for all titrations performed.
2. Also include the concentration of the NaOH solution from each titration and an
uncertainty analysis for each run. Remember that the error for a net weight of KHP is
0.0010 g and the error for a net volume of NaOH is 0.10 mL. You need to write out,
in detail, one example of how you calculated the concentration, and include the
calculations of the upper and lower limits of the concentration.
3. If you do have agreement between two runs, the final concentration is the average of
the two runs. Also report the error limits for the average. Average the upper limits of
the two runs to obtain the upper limit of the average. Likewise average the lower limits
to obtain the lower limit of the average. Again, do not average two runs that do not
overlap.
4. Report the molecular weight for your Unknown Acid. Make a table containing the
net weights and net volumes for all the titration. Write out one example calculation of
the molecular weight. You do not need to perform an error analysis for this part.
Remember to include the Unknown number of your acid.
20
21
sodium is also present. This test takes only 5 or 10 min and can provide useful
information.
Part B: Analysis for Water of Crystallization
A certain number of water molecules are needed to hold together the mineral crystal.
They are weakly bonded to the ions, and heating or sometimes simply exposure to a
dry atmosphere causes them to be lost. Usually the water of hydration has a definite
stoichiometric ratio to the ions in the salt. This loose bond between the water and the
rest of the ions is signified by the dot as, for example, in CaSO4 2 H2O. This is the
formula for gypsum and states that the crystal contains two water molecules for each
Ca2+ or SO42- ion. Often the color and hardness of the hydrated form of the crystal and
the anhydrous (water-free) form are strikingly different. A practical use for the
hydration phenomenon is the setting of plaster of Paris CaSO4 H2O. When mixed
with water, it slowly turns into solid gypsum.
How to Measure the Water of Hydration
You will do the reverse of the reaction just described in the plaster of Paris example.
You will take a hydrate and heat it until it is anhydrous.
MSO4 x H2 O MSO4 + x H2 O
1. Heat a small crucible (not a filter crucible) for about 2 min on a clay triangle to dry it
thoroughly. When it is cool, determine the precise weight of the crucible and cover.
to handle the hot crucible.
Use crucible tongs
2. Record the sample unknown number immediately upon obtaining the sample.
Weigh the sealed vial, transfer between 0.5000 and 2.0000g of mineral to the crucible,
and reweigh the vial to find the exact weight transferred.
3. Place the crucible on the triangle, and put on the top, so that it does not completely
cover the crucible but sits jauntily like a beret. Heat gently at first, and keep the flame
moving to avoid hot spots. Increase the heat moderately but do not permit the crucible to
become red hot. Overheating may cause the salt to decompose. Using tongs,
occasionally remove the cover to see if the dehydration appears to be complete.
Observe any change in the appearance of the powder caused by the loss of the water
molecules. When the dehydration is complete (15 min), continue to heat for another
five min.
4. Check your desiccator. This is a device to keep objects free from atmospheric
moisture, and depends on a chemical drying agent, CaCl2. Is the CaCl2 drying agent
caked or moist? If so, it should be replaced. If the seal is not tight, a little grease will
help.
22
5. Allow the crucible and contents to cool in the desiccator to prevent stray moisture
from being picked up. Weigh the sample accurately when cool, then reheat for 10 min
and see if a further decrease in weight occurs.
6. Repeat until the weight is the same, within uncertainty limits. Then you may be
certain that all the moisture has been driven off. This procedure is called drying to
constant weight. Remember that objects seem lighter when they are hot because of air
currents near the balance pan. Be sure to wipe off any stray drying agent clinging to
the outside of the crucible before weighing.
Strictly speaking, we have not shown that the only chemical change is loss of water.
This might be hard to prove. One could condense the water vapor and show that the
weight of water is equal to the loss of sample weight. Or, one could show that if water
is added to the anhydrous salt, the product is chemically identical to the original
mineral.
How to Do the Calculations for This Part
The calculations in this section exactly parallel the coal example in mathematical
operations section. Make a table showing the net weight of wet mineral and of dry
mineral, the net weight loss, and the grams of H2O per gram of mineral (Gwater), and the
moles of water per gram of mineral (Mwater). These values are calculated by
Gwater =
wt. water
wt. sample
M water =
moles water
Gwater
=
wt. sample 18.015 g/mol
where wt. water is the weight lost by the sample, and wt. sample is the weight of the
wet mineral sample at the start of the analysis. The value for the molecular weight of
water is 18.015 g/mol.
Show all the worst cases, and compute the range in the final number Gwater and Mwater.
Only the net weight of water lost and wet mineral sample have errors associated with
them.
Part C: Sulfate Analysis
We need to know how many grams of each gram of mineral are sulfate. Clearly, it is
not possible to separate the SO42- ion and weigh it.
In this situation, chemists use a technique called gravimetric analysis. One forms an
insoluble compound with the ion in question and collects the precipitate from the
solution. Your analysis will exploit the fact that BaSO4 precipitates out until virtually
all the sulfate ion has been removed from the solution. Then it is a simple matter to
filter out the solid BaSO4 and weigh it. The number of moles of BaSO4 formed will
equal the number of moles of SO4 2- present. The balanced equation is shown below.
23
Ba 2+ (aq) + 2 Cl - (aq) + M2+ (aq) + SO24 - (aq) BaSO4 (s) + M2+ (aq) + 2 Cl - (aq)
Note that the BaCl2 totally dissociates to the Ba2+ and Cl- ions, and no harm is done if an
excess of Ba2+ ions are added to the solution.
A gravimetric analysis can be made very accurate because the analytical balance if
potentially one of the most accurate instruments we have available. An accurate
weight of BaSO4 is all that is needed to calculate accurately the number of moles of
sulfate in the original mineral sample.
CAUTION:
Concentrated Hydrochloric Acid, HCl, is corrosive and must not be allowed to touch
the skin, eyes or clothes. Clean up any spills, and rinse skin with water. Inform
your TA of any spills.
How to Do the Sulfate Analysis
1. Weigh the original sealed vial containing your original mineral sample. Transfer
about 0.2000 to 0.5000 g to a clean but not necessarily dry 250 mL beaker. Reseal the
vial and reweigh to find the amount of sample transferred. (If you go modestly over
0.7 g, you can still save the experiment by adding proportionately extra BaCl2 . More
than 0.7 g will yield so much product that filtering will be very slow. Clean out the
beaker and begin again.) If your sulfate mineral is only marginally soluble in water,
you may find that even 0.2 g is too much and that you need much smaller samples.
2. To the beaker add 100 mL of deionized water and 3 - 5 drops of concentrated HCl.
Bring to the boiling point, then using beaker tongs remove the burner so that the liquid
stops boiling. Obtain 10 mL of BaCl2 solution in your graduated cylinder. Once all of
your compound has dissolved, add a drop of 0.5 M BaCl2 solution, and swirl. Then add
the remaining 10 mL of BaCl2 solution. This method creates larger and more easily
filterable crystals of BaSO4. Cover the beaker with a watch glass and boil gently for at
least 5 min. Then let the beaker stand for a few minutes. The boiling process enables
the small crystals of BaSO4 to dissolve and larger ones to form, a process called
digestion.
3. While you are waiting for the digestion process to take place, examine a filter
crucible. Check to see if they have cracks in the fritted bottom by holding them up to
the light. If it is necessary to clean them, put them in the oven to dry for 1 hr afterward.
Do not weigh with a wet frit! Weigh a clean, dry filter crucibles. When the digestion
process is completed, filter the suspensions of BaSO4 through the crucible. Your
teaching assistant will demonstrate the best method. Speed up the filtration by
allowing the solid to settle and pouring the supernatant liquid through the filter first
(see figure below).
24
Use a rubber policeman to scrub the inside of the beaker, and use the jet of water from a
wash bottle to direct the solid into the crucible. You must transfer ALL of the solid to
the filter crucible, and if any is spilled, discard the sample. Run about 5 mL of water
through the filter, crucible to "wash" the sample, that is, to remove any ions clinging to
the solid. Your teaching assistant will show you how to use the aspirator to dry the
crucible. Put your crucible in a 100 mL beaker, and give your teaching assistant the
beaker with a scrap of paper inside that has your name on it. The sample will be oven
dried to eliminate the last traces of water. Then you will weigh the crucible (when cool)
to determine the weight of BaSO4 .
One can do a gravimetric analysis only when no interfering ions are present. In this
precipitation, acid (HCl) is added to destroy any carbonate ion (by converting it to
carbon dioxide (CO2), which will bubble off) and any hydroxide ion (by converting it to
water). Both barium hydroxide (Ba(OH)2) and barium carbonate (BaCO3) are insoluble
in water and would be collected on the filter with the BaSO4 desired. Thus, hydroxide
and carbonate are "interfering" ions for this analysis. The interfering ions are different
for each type of precipitation. Systematic errors result whenever the reagent
precipitating the ion you wish to measure precipitates stray ions as well.
How to Do the Calculations for this Part
The calculation relies on the known relationship between the weight of BaSO4 and the
amount of sulfate in the sample. From the balanced equation, we see that the BaSO4
and SO42- have a stoichiometric ratio of 1:1.
25
1. Convert the weight of BaSO4 to the number of moles of BaSO4 by using the molecular
weight (233.4 g/mol). Then note that the number of grams of SO42- in this material is
the number of moles of BaSO4 multiplied by 96.06 g/mol. This is also the number of
grams of SO42- in the original mineral sample. Divide the weight of SO42- by the weight
of sample used in this part of the analysis to get the fraction of sulfate, Gsulfate.
moles sulfate
Gsulfate
Msulfate =
=
wt. sample
96.06 g/mol
3. Calculate the upper and lower limits for the values of Gsulfate and Msulfate. Only the net
weight of BaSO4 and mineral sample have errors associated with them.
26
There are two types of resins. Cation exchange resins are materials that contain many
SOOH or COOH groups that can exchange the cation H+ for other positive ions.
Anion exchange resins have groups such as -N(CH)3+OH- or -N(CH3)+Cl- that exchange
OH- or Cl- anions for other negative ions. Each location where exchange can occur is
called an exchange site. One long chain molecule may have many exchange sites.
When one ion is exchanged for another, electrical neutrality must be preserved. Ion
exchange resins work by substituting one type of cation (anion) for an electrically
equivalent quantity of another cation (anion). For example, if a table salt (NaCl)
solution is passed through a cation exchange column, HCl emerges from the column
since the Na+ ions are replaced by H+ ions. Conversely, NaOH emerges if NaCl is
passed through an anion exchange column because OH ions replace the Cl- anions.
(Water is "deionized" by passing it first through a cation exchange column and then
through an anion exchange column. The first converts NaCl to HCl and the second
changes HCl into HOH, water.)
Figure 3.2 Ion Exchange Column
SO3 H
SO3 H
SO3 H
+ H+
+ M2+
SO3 M+
before exchange
after exchange
27
CAUTION:
Sodium Hydroxide, NaOH, and Hydrochloric Acid, HCl, are corrosive and must not
be allowed to touch the skin, eyes or clothes. Clean up any spills, and rinse skin
with water. Inform your TA of any spills.
How to Run the Ion exchange on the Mineral
You will be loaned an ion-exchange column containing resin. Throughout this
experiment the column must not be allowed to run dry, because this creates channels
through which solution can run without being exposed to the resin.
The ion-exchange column should be filled with H+, although it is best to convert it
again, in case the previous user has been sloppy. Check to see that there are no air
bubbles in the resin before beginning. Drain the liquid until it is within 0.5 cm of the
top of the column. Add about 15 mL of 3M HCl to the column, and then allow it to
drain slowly. Continue to add HCl to the column until 100 mL have passed through it.
Then wash the resin with at least 100 mL of deionized water. After the rinsing, collect a
drop of the water emerging from the column (the eluate.) This drop should become
only faintly cloudy when a drop of silver nitrate solution is added. We have now filled
all of the exchange sites on the resin with H+ and removed all excess acid.
Weigh the vial containing the mineral and transfer a new sample of not more than 0.3 g
to a clean, but not necessarily dry, beaker. Reweigh the vial to find the exact amount of
mineral that was transferred. Add about 40 mL of water to the mineral, and add all of
the resulting solution to the column. Collect the eluate in a clean conical flask. Rinse
the beaker with a wash bottle and add the rinsing to the column. Also run 100 mL of
distilled water through the column, and collect in the same conical flask. Run all
substances through the column slowly. (How slow is slow? You should be able to count
the drops.) Keep the eluate in a conical flask, and store until you are ready to titrate. (If
you need to run a second sample through the column, separately collecting the eluates.
It is not necessary to regenerate the column between samples.)
How to Analyze the Eluate
The eluate now contains as many H+ as there were positive charges in the mineral
sample. According to the equation below
NaOH (aq) + H+ (aq) H2 O (aq) + Na + (aq)
each mole of NaOH will react with exactly 1 mol of H+. Because each mole of doublepositive metal ions in the mineral will produce two moles of H+ ions, we can easily
moles of metal ions from the number of moles of NaOH consumed.
calculate the
You have already determined the concentration of your NaOH solution by allowing it
to react with a known weight of KHP. Titrate the eluate with this standardized
solution of NaOH.
28
If you overshoot the end point, all is not lost! Add a known amount of solid KHP, and
then titrate carefully to the end point. The bookkeeping may be a bit more difficult, but
it sure beats starting over!
Doing Calculations and Uncertainty Analysis
The calculation here is not mysterious. It is really very similar to the calculations done
in the standardization of NaOH.
1. You should have computed the concentration of your NaOH solution and its upper
and lower limits when you were standardizing the solution.
2. Compute the moles of H+ in the eluate sample from the moles of NaOH required for
neutralization. Remember to correct for any back titration. Compute the number of
moles of metal ion in the sample you used; remember that the ion exchange column
releases two H+ ions for each M2+ ion exchanged. Then calculate Mmetal the number of
moles of metal ion per gram of mineral.
M metal =
3. Compute upper and lower limits for the number of moles of metal ion in your
sample by following the same procedure as that in Step 2. The upper limit of Mmetal will
be theupper limit of the number of moles of metal ion divided by the lower limit of the
weight of mineral sample used. With this hint, you should also be able to work out the
lower limit.
For the Report
Now you are ready to work out the formula of the mineral.
1. To obtain the formula, compute the following, if you have not already done so.
Mwater = the number of moles of water per gram of mineral
Msulfate = the number of moles of sulfate per gram of mineral
Mmetal = the number of moles of dipositive metal ion per gram of mineral*-+.
Make a table containing the upper and lower limits for all your M and G values.
2. Charge Balance: The correct formula must have an equal number of positive and
negative charges. If the experiment has been carried out correctly, the limits for Msulfate
and Mmetal should overlap. Choose one best value, since it is clear that Msulfate must
equal Mmetal by charge balance.
29
Gmetal
M metal
What are the upper and lower limits of the molecular weight of the metal ion?
30
Apparent Atomic
Weight of M2+ (g/mol)
24.31
24.31
24.31
45.96
45.96
K2Mg(SO4)24 H2O
K2Mg(SO4)26 H2O
Na2Mg(SO4)22 H2O
KAl(SO4)212 H2O
51.28
51.28
35.13
33.04
31
CH2COOH
H3C
HO
CH2COOH
H2C
CH2
HOOCH2C
C
CH
HC
C
C
H
C
H
C
HC
HC
CH3
SO3H
CH
C
H
In this structural formula, only one of the many possible resonance forms is shown.
The " systems of all three rings overlap to form a continuous delocalized system, and
the electrons can migrate allover this network. This gives rise to unoccupied low-lying
electronic states and to the absorption of visible light. The compound can also undergo
significant rearrangements in acidic or basic media. (It has structural elements similar
to those of phenophthalein.)
The protons of the four -COOH groups are easily lost, and so we will think of xylenol
orange as a tetraprotic acid and abbreviate it H4Q.
What is the reaction you will Study?
H4Q (aq) + Al3+ (aq)
Because the only colored species in this reaction are the QAl complex and the H4Q, you
should be able to follow the reaction by monitoring one of them. You will do this by
measuring the intensity of the color of the QAl complex.
32
The intensity of color is related to the concentration by Beer's Law. This relationship
and the use of the Spectronic 20, which is the instrument used in this experiment, are
discussed in the first section of Instrumentation.
Measuring the Equilibrium Constant
To measure the equilibrium for the above reaction we need to be able to determine the
concentration of the QAl molecule. Luckily, the H4Q is not the same color as the QAl,
so we will make our observations at a wavelength of 550 nm, which is close to the
maximum absorbance of the QAl molecule. Nevertheless, there will be some
absorbance due to the unreacted xylenol orange, and a "blank" or control must be used
to overcome this.
1. Clean three test tubes of 1 cm diameter. Pipette exactly 2.0 mL of xylenol orange into
each. The easiest way to provide suction for the small pipet required is to use a syringe
and to connect it with a small section of rubber tubing. After the xylenol orange, pipet
exactly 2.0 mL of Al3+ into the three test tubes. One of these test tubes will remain at
room temperature, at which no reaction takes place, and will be a control. The other
two test tubes will be heated in a water bath.
2. Add about 150 mL of water to a 250 mL beaker, immerse two of the test tubes, and
bring the water to the boil. Continue to boil the water for at least 5 min to allow the
equilibrium to be established. Then use tongs to remove one of the test tubes and place
it in an ice bath. Using the tongs, place the second test tube in a test tube rack. Use the
beaker tongs to pour off the hot water into a thermocup (put one Styrofoam cup inside
another). Still using tongs, transfer the test tube from the rack to the thermocup, which
will slow the cooling process. Use a thermometer to measure the water temperature.
When the water cools to about 90 85 C, make the first measurement. Then repeat
every 5C or so, until the sample reaches 50C.
3. To make a measurement, put your control test tube of xylenol orange into the
Spectronic 20 and adjust for 100% transmittance. Pull out the test tube in the hot water
bath, noting the temperature. Wipe it dry with a paper towel, and put it into the
Spectronic 20 sample holder. Read the % transmittance of the mixture, and then replace the test tube into the water bath. Be careful not to bum yourself or to spill the
solution into the Spectronic 20. Perform this operation quickly to avoid the solution
cooling while in the Spectronic 20.
At the end of the experiment you should have a series of measurements of the %
transmittance for at least six temperatures. You can repeat the experiment any number
of times by reheating the contents of the tube to 100 C, waiting 5 min, and letting the
tube cool slowly. Be sure to record the concentrations of the solutions used and all the
other label information.
4. The last and, in many ways, the most interesting experiment measure the %
transmittance of the test tube that has cooled in the ice bath.
33
" % transmittance %
A = log10 $
'
#
&
100
The equilibrium concentrations of QAl are computed from Beer's Law:
=
The extinction coefficient, , is 25,000 L/mol cm for this compound (a very intense
color, comparable to that of permanganate). Take the path length as 0.9 cm.
2. The concentration of H+ is from the pH marked on the container and will be virtually
unchanged throughout the reaction.
3. The concentration of the other two species can be obtained by means of
stoichiometry:
!"Al3+ #$ = !"Al3+ #$
!"QAl- #$
and [ H 4Q ]equil = [ H 4Q ]initial !"QAl- #$
equil
initial
equil
equil
Remember that the initial concentrations are referring to the starting reagent
concentrations in the test tubes. These concentrations are half those marked on the
bottles because of dilution.
3. From these equilibrium concentrations, compute the equilibrium constant (K) at each
temperature.
4
!"QAl- #$!"H + #$
K=
[ H 4Q]!"Al3+ #$
4. The fact that the equilibrium constant depends on temperature is useful, because it
allows you to measure values of G, H, and S for the reaction. You can easily
obtain the equilibrium constant from the concentrations. You know that
G = RT ln K
34
ln K =
H # 1 & S
% (
R $T' R
35
36
5 C2O42- + 2 MnO4 + 16 H+
This reaction can be used for titration only if the solution temperature is at least 70 C
to 90 C.
Because this exam was run as a contest for many years, the most common causes of
failure seem to be
1. Failing to understand the instructions.
2. Preparing solutions poorly, or "mistreating" them. This category breaks down into a
number of subproblems, such as not mixing the solutions thoroughly. The average
student seriously underestimates the shaking required to mix things in a volumetric
flask. The solutions must be absolutely uniform it is impossible to shake them too
much.
Contaminating the primary standard. This can happen by placing the solution in a
container that is wet on the inside, because the beads of water can cause significant
dilution. Primary standard solution should not be poured back from the buret into the
storage bottle (this practice is dangerous and introduces contamination or stray water).
Adding the primary standard to a buret or funnel containing stray beads of water can
also introduce contamination.
3. Not allowing the primary standard solution to cool before diluting to the mark (the
H2SO4 solvent contracts as it cools).
Mixing up records. (This is inexcusable, but it keeps happening. Make sure it does not
happen to you.)
4. Making blunders in the use of, for example, the buret and the balance. The practical
exam is extremely efficient at finding people who, for example, do not know how to
read the balance correctly. If you have any lingering doubts, ask your teaching
assistant, even if you think you really ought to know.
Averaging good runs with those in which blunders occurred. Averaging does not
reduce systematic errors; it only means that good data are spoiled. Only average your
results if you are reasonably sure that no blunders occurred that could affect them. It is
far better to have one good standardization and one good titration of the unknown than
to have three poor runs.
37
38
with the KMnO4. The reverse procedure does not work as well; compute the ratio of
volumes used.
SAVE BOTH SOLUTIONS FOR THE EXAM
4. On the day before the practical exam, reread the section of the book called How to
Be Successful in Lab so that you recall all of the special tricks of titration.
Doing the Oxalate Analysis
You will have 2 hrs. and 30 min to do this part.
1. Obtain a sample from the stockroom. A signed receipt is required.
2. Because it gradually deteriorates, the KMnO4 solution must be standardized on the
day it is to be used. Shake the solution vigorously for one full minute before beginning.
Standardize the solution against oxalate in the way described in Step 3 in Preparing the
Solution, above. Do not forget to heat the solution.
3. For the titration of the unknown, place about 1.0000 g of the unknown solid in a
conical flask, recording the exact weight. Weigh out one sample at a time, so that if too
much or too little KMnO4 is used, you can adjust the next sample size to be more
convenient. Dissolve the unknown in at least 25 mL of 3M H2SO4. You may wish to
heat it to speed up dissolution. Remember, the reaction occurs rapidly only when the
solution is hot, so you will have to heat until the temperature of the solution is 70C to
90C before titrating.
4. Ideally, you should do three standardizations and three titrations. The ratio of
KMnO4 to Na2C2O4 should not be drastically different from that on the dry run day.
Work with deliberate speed and do not waste time, but it is much preferred to obtain
one good pair of runs than three poor ones.
5. When "time" is called, stop work and complete the calculations on the report form.
You may choose the mean, the median, or the best value, or you may just guess. If your
answer for oxalate content is not between 10% and 40%, you have probably made a
mathematical error. Your grade will be based solely on how close your number is to
the correct value of the unknown. Give some thought to the choice of mean, median, or
best value.
6. Avoid the following errors:
a) Mixing up burets.
b) Forgetting to heat the solution (end point appears sluggish).
c) Forgetting to add H2SO4 where needed (end point brownish).
7. Before you leave, be sure to clean your work area, wiping up any spills. You will
dispose of the solutions during the Check-out period. Keep the sample vials until then.
39
The instrument is now correctly calibrated, but for one particular wavelength only. To
take a reading, place a cuvette containing your sample in the sample holder, close the
cover, and record the transmittance after the needle has settled down. The cuvette
must be free of fingerprints, bubbles, and other dirt.
When you are using the Spectronic 20, keep the test tube or cuvette clean; avoid leaving
fingerprints on the bottom part where the light passes through. There should be no air
bubbles on the inside, and solutions with suspended material will result in odd
readings.
The cuvette is cylindrical and therefore acts as a lens, focusing the light to some extent.
The cuvette should therefore be lined up in the same way each time. There is a mark on
40
some cuvettes, which can be lined up with a line on the cell holder. If there is no mark,
make a small one at the top of the tube using a wax pencil.
The calibration with the blank must be redone at each new wavelength and should be
repeated every few minutes, even at the same wavelength, to compensate for any drift
in the electronics.
Remove the cells in order to fill them. Water and electricity do not mix!
How the Spectronic 20 Works
You will be measuring the amount of light absorbed at a chosen wavelength. The
Spectronic 20 is a typical absorption spectrophotometer, and it has the following basic
parts:
1. A source of light. For the Spectronic 20, this is a tungsten lamp. White light
comes out of the bulb, and lenses focus it into a narrow, parallel beam.
2. A monochromator. This is a device used to break up the light into several
wavelengths and to block off all but the wavelength of interest. In your
instrument, the light is broken up by a diffraction grating and sorted by an exit
slit, which stops all but one wavelength band.
3. A cell holder to hold a cuvette or sample test tube. In the Spectronic 20 there is an
occluder to prevent light from passing through if there is no sample in the holder.
4. A photocell or other device used to "see" the light and to give a numerical value to its
intensity. The phototube will measure all the light remaining after the sample
has done the absorbing.
All spectrophotometers have the same basic parts, but obviously different sources of
radiation and types of receptors are needed for different regions of the electromagnetic
spectrum. One of the reasons for taking time to understand the Spectronic 20 is that
once this instrument is understood, mastery of more complicated but basically similar
instruments is easy.
How the Spectronic 20 Controls Do Their Job
The wavelength controller adjusts the position of the diffraction grating so that different
colors of light are selected by the slit.
The zero control adjusts the electronics. With the sample compartment cover down and
no sample, this should be adjusted until the meter reads 0 for transmittance. This
control adjusts for the so-called dark current, the current transmitted by the photocell in
the dark.
The 100 percent T control, a knob on the right, controls the amount of light output. A
mechanical linkage is used to drive a wedge into the light beam. This should be
adjusted so that with a test tube full of solvent in the sample holder, the transmittance is
41
100 percent. This allows the operator to correct for any stray absorbance resulting from
the solvent.
COLOR AND SPECTROSCOPY
When various wavelengths of light enter the eye, we see different colors. The rainbow
is an illustration of a spectrum, in which white light is sorted into its various
component wavelengths, each of which appears as a different color. Such an effect can
also be produced by a prism or diffraction grating.
The range of wavelengths visible as colors for humans is about 700 nm (nanometers,
where 1 nm = 10-9 m) to about 380 nm. It is interesting that some animals can see colors
in the wavelength region beyond violet (ultraviolet) or in the region beyond the red end
of the spectrum (infrared). Humans have to rely on sophisticated instruments to make
observations in these regions of the electromagnetic spectrum.
The color of light transmitted by a solution or reflected by a surface is the light that
remains after some colors have been preferentially absorbed. Chemists are interested in
the region at which light is absorbed, because this can supply information about the
structure of the molecule.
We therefore need to know what wavelength is absorbed most preferentially and also
the intensity of the absorption. The wavelength of absorption is therefore related to the
energy of the light being absorbed by the molecule. Thus, the color of the absorbed
light provides direct information about the kind of molecular transition or the kind of
molecule doing the absorbing. The intensity of absorption is related partly to the type
of molecule, but it also depends on other variables. This section first looks at the color
alone.
The Relationship of Color and Wavelength
In deciding what color something is, you have to distinguish between objects that emit
light and objects that reflect (or transmit) light. A Bunsen burner flame is blue because
the combustion reaction products emit blue light. For such objects, the relationship
between color and wavelength is the same as for sunlight dispersed by a prism. The
solutions you will use are actually involved in the reverse process; instead of emitting
light of a certain wavelength, , they absorb light at wavelength, , and transmit the rest
of the light. So a solution of CuSO4 held up to a light appears blue because the Cu2+ ion
absorbs yellow light and transmits the blue light, which is not absorbed. To figure out
what color light is absorbed if you know what color is transmitted (complementary
colors) use Table 5.1.
The experiment you will do uses solutions that absorb light rather than emit light. The
of the absorbed light is the wavelength of interest to chemists, because it can provide
clues about molecular structure.
42
Our instrument, the Spectronic 20, is calibrated in nanometers (nm), which are
wavelength units. However, the wavelength may also be expressed in Angstrom units
(), which are equal to 10-10 m. Also, infrared workers frequently use cm1 (the
reciprocal of wavelength). This latter unit can be thought of as wave crests per cm and is
really an energy unit.
TABLE 5.1: Relationship Between the Color Seen and the Wavelength of Light
Absorbed Light Absorbed Color You See
Light Absorbed
Wavelength (nm)
400-440
440-480
480-490
490-500
500-560
560-580
580-600
600-610
610-750
Color
violet
blue
green-blue
blue-green
green
yellow-green
yellow
orange
red
43
If the concentration is in stated moles per liter and the path length is in centimeters, the
extinction coefficient will be in liters per mole-centimeter.
The Units of Color Intensity
The meter or chart recorder supplies the intensity of absorption at a given wavelength.
It is calibrated in either or both of the two following ways:
1. Transmittance. This is the fraction of the initial number of photons that pass
through the sample without being absorbed. (Sometimes the transmittance is
expressed as a percentage.)
2. Absorbance. This is the negative logarithm of the transmittance (expressing the
latter as a fraction). It is unitless. Absorbance and optical density (OD) as the
biochemists call it) are the same thing.
The relationship between absorbance A and transmittance T is
A = -log10 T
where T is expressed as a fraction. The following example makes this clearer:
EXAMPLE: A student records a transmittance of 30 percent. What will be the
transmittance if the concentration of the solution is doubled?
Solution: Express 30 percent as a fraction, or 0.30, and compute the corresponding
absorbance,
A = -log(0.30) = 0.52
If the concentration is doubled, the absorbance is doubled (from Beer's' Law). Hence,
the new solution will have an absorbance of 1.04, and a transmittance of 9 percent.
44
Mathematical Operations
This part is largely concerned with uncertainty calculations. You will need to master
the first section to complete most of the lab reports, but the second and third sections
may well be assigned by your professor to deepen your understanding of this topic and
its applications.
If you have had to struggle with uncertainty in a previous lab course, fear not. The
method presented here is a lot simpler, and it works well enough for most student lab
work (and for a good deal of real scientific work as well.) Students who had trouble
with traditional methods can easily cope with uncertainty when it is presented in this
new way, so take heart!
A. CALCULATING UNCERTAINTY BY THE WORST CASE METHOD
There are two sorts of problems with measurements. The first kind is that of systematic
error, the situation in which one is measuring not quite what one supposes, because of,
for example, defects in instruments or technique. Such systematic errors can be
controlled only by improving the design of experiments.
This section discusses the other sort of ambiguity in data, the intrinsic uncertainty due to
the limitations of the instruments. It is assumed here that the desired quantity is in fact
being measured. However, the question is asked: how reproducible would the
measurement be if it were repeated in the same way many times? This reproducibility
or precision is of major importance in planning experiments.
Systematic errors and uncertainties differ in one crucial way. There is no way that
systematic errors can average out. If, for example, a sample from a gravimetric
experiment is not sufficiently dried before weighing, the student can weigh it one
hundred times and the problem will not be cured. However, if a sample is correctly
prepared, more weighings averaged will produce a result of greater precision.
A beginning student has far too much faith in averaging. If the measurements being
averaged contain repetitions of the same systematic errors, the result cannot be purified
by running it through the calculator!
Many classic textbooks draw a distinction between precision (the degree of clustering)
and accuracy (the absence of systematic error). Look at Fig. 6.1 showing a bear target
being shot by a marksman.
45
Computing Uncertainties
Every measurement is uncertain in that it probably cannot be exactly reproduced a
second time. Uncertainty is a technical term for the reproducibility of the measurement.
If, for example, you weigh a copper penny on a balance five times, you will probably
observe some variation in the last decimal place. Such a sequence of results might be
3.0110 g, 3.0114 g, 3.0120 g, 3.0111 g, and 3.0115 g. The average value is about 3.0115 g,
but the range of values is at least 0.0005 g from that value. The number 0.0005 is
called the absolute uncertainty of the measurement. Clearly, the real value (whatever
that is) is somewhere between 3.0110 g and 3.0120 g. These two values are referred to
as worst cases or the upper and lower limits.
Similarly, for any other instrument there is a range of absolute uncertainty in a
measurement related to its observed reproducibility. Some are given below, for the
most common models of instruments in student labs.
buret
balance
360 thermometer
The National Bureau of Standards (NBS) sets the following standards for absolute error
tolerable in Class A glassware (Tables 6.1 and 6.2):
When you are confronted with a new instrument and need uncertainty information, it
is sensible to try repeating the same measurement until some idea of the reproducibility
is evident. Then it is easy to choose reasonable values for worst cases. (There is, of
course, some judgment involved.)
46
Tolerance Common
Tolerance Class A
10 mL
0.04 mL
0.02 mL
50 mL
0.10 mL
0.05 mL
100 mL
0.16 m
0.08 mL
250 mL
0.24 mL
0.12 mL
500 mL
0.30 mL
0.15 mL
Tolerance, Transfer
Tolerance, Measuring
1 mL
0.006 mL
0.01 mL
2 mL
0.006 mL
0.01 mL
5 mL
0.01m
0.02 mL
10 mL
0.02 mL
0.03 mL
25 mL
0.03 mL
0.05 mL
47
Upper Limit
20.7995g
20.8000g
20.8005g
19.9995g
20.0000g
20.000g
0.7990g
0.8000g
0.8010 g
These upper and lower limits are calculated in exactly the same way as in the buret
example:
Upper limit: 20.8005 g 19.9995 g = 0.8010 g
Lower limit: 20.7995 g 20.0005 g = 0.7990 g
So the net weight of wet coal is 0.8000 0.0010 g.
48
weight loss
100
original weight
Hence, the true percentage of water lost lies between 10.14 percent and 9.86 percent.
This problem could be simplified by realizing that the "net" quantity rule (the corollary,
Rule 4) applies to Steps 1 and 2.
How to Calculate Uncertainty in a Concentration
A solution is made up in an l00 mL volumetric flask. The substance has a molecular
weight of 120.0 g/mol, and 2.6754 g are weighed out. What is the uncertainty in the
concentration?
Concentration =
2.6754 g
(0.10000 L)(120.0
g = 0.2229 !
)
mol
The weight is determined by weighing a vial containing the solid, then transferring the
solid quantitatively to the flask, and reweighing the vial. The absolute uncertainty in
each weighing is 0.0005 g. The weight is a net weight, so the absolute uncertainty of
the net weight is 0.0010 g (by Rule 4).
49
The flask is 100.00 mL nominally and has an absolute uncertainty of 0.08 mL (if it is
Class A). The volume limits, therefore, are 99.92 mL and 100.08 mL. Combining the
upper limit of weight with the lower limit of volume provides the upper limit of
concentration.
Upper limit =
Lower limit =
2.6764 g
(0.09992 L)(120.0
2.6744 g
(0.10008 L)(120.0
g = 0.2232 !
)
mol
g = 0.2227 !
)
mol
The last step of this example illustrates the division rule about division of worst cases.
Problems for You to Solve
1. Jo Superior is a student at a college where the balances are accurate only to 0.005 g.
Into a crucible that weighs 20.100 g empty, she puts some copper powder and reweighs
it, obtaining a reading of 21.100 g. An excess of sulfur is added, and then the mixture is
strongly heated under a hood. The excess sulfur vaporizes, leaving a nonvolatile
compound of copper and sulfur in the crucible. The weight of the crucible and
compound is 21.554 g. The atomic weight of copper is 63.54g/mol; atomic weight of
sulfur is 32.06 g/mol.
Compute the upper and lower limits for the ratio of moles of sulfur to moles of copper.
2. A titration of 0.2686 g of an unknown acid HX requires 23.06 mL of NaOH solution.
The concentration limits of the base solution are 0.0872 M and 0.0884 M. What are the
upper and lower limits for the molecular weight of the HX?
3. A l0 mL Class A pipette is used to deliver a sample of solution of unknown acid HX,
which is titrated with 23.06 mL of NaOH, the concentration of which can be taken as
exactly 0. 01000 M. What are the upper and lower limits for the concentration of the HX
solution?
50
Given the formula above, it is easy to compute relative uncertainty in any specific case.
Note that whereas the absolute uncertainty depends on the instrument, the relative
uncertainty depends on the size of a measurement. For example, if you weigh 1.0000 g
by difference, our absolute uncertainty will be 0.0010 g. This absolute uncertainty is
the same if you measure 0.1000 g. However, the relative uncertainty is different:
for 1.0 g !" =
0.0010 g
100 = 0.1 %
1.0000 g
0.0010 g
100 = 1.0 %
0.1000 g
51
Then, if you multiply Y itself by the relative uncertainty of Y, you obtain the absolute
uncertainty of Y and can construct worst case upper and lower limits. In summary,
then, to perform this trick do the following:
1. Convert absolute uncertainties to relative uncertainties for each of the
measurements, to be multiplied or divided.
2. Add all of the relative uncertainties.
3. Compute the absolute uncertainty of the result by multiplying the relative
uncertainty of the result by the result itself.
4. Construct worst case limits for the result.
Understanding Why This Short Cut is Useful
Not only does this short cut save time, it pinpoints the weakest link in the chain of
measurements being used to lead to a conclusion, because this one will have the highest
relative uncertainty.
The result you obtain may not be exactly identical to that from the worst case method,
but it will probably differ only slightly. Because absolute uncertainties are often just
estimates anyway, you do not need to worry.
Remember the limitation of this trick: All measurements must be related to the final
derived value by multiplication or division, or by multiplication or division by a
constant. This short cut does not work for addition or subtraction.
An Example Uncertainty in a Standardization
A student finds that 0.5089 g of potassium acid phthalate exactly neutralizes 23.06 mL
of a NaOH solution of unknown concentration.
Step 1. Calculate the concentration itself. The number of moles of potassium acid
phthalate used is
0.5089 g
= 2.492 10!! mol
204.22 g/mol
Since each mole of this substance reacts with the same number of moles of NaOH,
there must be 2.492 10-3 mol of NaOH in 23.06 mL of solution. So the
concentration will be
2.492 10!! mol
mol
= 0.1080
0.02306 L
L
52
Step 2. Compute relative uncertainties. The potassium acid phthalate was weighed by
difference; hence, the uncertainty in each weighing is 0.0005 g, and the total
absolute uncertainty is 0.0010 g. Thus, the relative uncertainty of the weight of
potassium acid phthalate is
0.0010 g
100 = 0.20%
0.5089 g
This is also the relative uncertainty of the number of moles of potassium acid
phthalate, and, therefore, of the number of moles of NaOH (since we assume that
there is no uncertainty in the molecular weights).
The volume of NaOH is measured by difference. Because the initial buret reading is
uncertain by 0.05 mL and the final reading by 0.05 mL, the total absolute
uncertainty is 0.10 mL. This gives the relative uncertainty on the volume as
0.10 mL
= 0.43 %
23.06 mL
Step 3. Add the relative uncertainties. The relative uncertainty of the NaOH
concentrations will be given by
RU of NaOH concentration = RU of NaOH mol + RU of NaOH vol
RU of NaOH concentration = 0.20% + 0.43% = 0.63%
Step 4. Convert the RU of the result back to an absolute uncertainty. The absolute
uncertainty will clearly be
0.63%
100
0.1080 ! = 0.0007!
Step 5. Construct worst case limits. Thus, the true value of the NaOH lies between
0.1073M and 0.1087 (0.1080 M 0.0007 M).
53
54
implies precision to the last place, and 2.0 implies only that the number is between 1.9
and 2.1.
The digits in a number that are known with certainty, plus one more, are customarily
reported. These are known as significant figures. Thus, the number 2.03 10-2 implies
that the value is between 2.02 10-2 and 2.04 10-2 (if no uncertainty is explicitly stated).
Although there are formal systems for keeping track of significant figures, if you
understand what it means to be "significant," you will never fall into the trap of
reporting 8.6412 for a number that is somewhere between 8.63 and 8.65.
Units should be attached to all numbers that need them, especially final results.
How to Prepare Graphs
A number of lab reports require that you prepare graphs. Over the years, scientific
journals have developed rigid standards for graphs. Your work must conform to these
standards, so that you acquire good habits.
The standards are as follows:
1. The variable being measured (dependent variable) is on the y-axis (vertically).
The variable being adjusted by the experimenter (independent variable) is on the
x-axis. Axes should be labeled in capital letters, with the unit in small letters in
parentheses.
2. Points must be large enough to be seen, and different series of data may be
indicated only by shape and symbol, not by color. There must be a key, caption,
or title explaining what the graph means.
Do not connect the points with a line. Let the shape of the line reflect the underlying
relationship. A linear relationship calls for a straight line; curves may be appropriate if
the underlying relationship is more complicated. Computer generated graph are
acceptable and preferred.
How to Write the Discussion
Most students seem to breeze through the calculations but are stumped by the
discussion part of the lab report. This section contains a report of one experiment, and
a sample discussion, to provide an idea of the possibilities.
Reporting the Data and Calculations of a Sample Experiment
Students were asked to identify the metal in a cylindrical slug by measuring the volume
and weight, and computing the density. The following are the data of the world
famous A Student:
55
Do not worry at this point about how the uncertainty limits are derived; this is
explained in the chapter on mathematical operations.
Discussing the Data of the Sample Experiment
The student's discussion, covering the above data, appears as follows:
The measured density was 2.86 g/mL with the "true" answer in the range of 2 .82 to
2.91 g/mL. This density is closer to that of aluminum (2.6989 g/mL) than to any other
metallic element. However, the density of the aluminum is outside the range of
uncertainty, and we therefore conclude that
1. The sample is not pure aluminum (possibly an alloy of aluminum with a denser
material?)
or
2. There may be an unnoticed systematic error in the measurement. Since the weight
measurement was made on an analytical balance, this could be wrong only if the balance
were improperly calibrated, a remote possibility.
The error, therefore, probably lies in the volume measurement if the hypothesis of
systematic error is correct. The volume would have to appear too small. This could
occur, for example, if the act of dropping the slug into the buret splashed water up onto
the sides, where beads would cling, and thus the rise in the water level would be less than
expected. This was not observed by the experimenter, and the buret was clean enough so
that such beads of water ran down immediately.
Given the extreme simplicity of the experiment, Hypothesis 1 seems more likely.
56
57