Determination of Ca

2+
and Mg
2+
in Water
by EDTA Titration
What are Lewis Acid-Base Reactions?
In your study of acid-base chemistry, you learned of different models for describing the behavior of
acids and bases. In the Arrhenius model, acids are substances that increase the concentration of
hydronium ion (H
3
O
+
) in aqueous solutions, while bases increase the concentration of hydroxide ion (OH
-
) in aqueous solutions. In the Bronsted-Lowry model, acids are proton donors and bases are proton
acceptors. Finally, the Lewis model considers acids to be electron pair acceptors and bases to be
electron pair donors. In this lab, you will use a Lewis acid-base reaction to determine the concentration
of Ca
2+
and Mg
2+
in a water sample. These ions and can inhibit the action of soaps and cause
precipitates called limescale. Water with high concentrations of these ions is said to be hard.
Because, these ions have a positive charge and vacant orbitals, they are good electron pair acceptors
and therefore Lewis acids. They can accept electron pairs from a donor that has an unshared pair of
electrons (i.e., a Lewis base). When a Lewis base donates its electrons to a metal ion to form a complex
ion, it is called a ligand. An example of a Lewis acid-base reaction involving the cyanide ion as a ligand is
shown below.


The reaction of the silver ion with the cyanide ion produces a complex ion in a process called
complexation. Complex ions are similar to polyatomic ions in that they are groups of covalently bonded
atoms that carry an overall charge. Complexation reactions are reversible and are characterized by an
equilibrium constant called a formation constant, K
f
. Values of K
f
are generally large, which indicates
that these equilibria strongly favor products, leaving a relatively low concentration of free metal ions in
solution. In fact, complexation reactions are commonly used to remove unwanted metal ions from
solutions. For example, EDTA is a complexing agent commonly added to food packaging in order to
complex metal ions that catalyze reactions that cause food to spoil.

What is EDTA?
EDTA is an example of a multidentate (many-toothed) ligand, which can bind metal ions through
multiple atoms. Multidentate ligands are also called chelates, which comes from a Greek word meaning
claw. Multidentate ligands, or chelating agents, wrap themselves around metal ions like a claw. The
figure below shows EDTA chelating a metal ion.

EDTA Chelating a metal ion


A more complete structure and a shorthand line structure of EDTA are shown below.

A line structure is a shorthand way of depicting an organic molecule. The lines in these structures
represent covalent bonds. Because all organic compounds contain carbon and hydrogen, these atoms
are not shown. A carbon atom is assumed to be present at the intersection of two lines. Because
carbon typically forms four bonds, the number of hydrogen atoms bonded to each carbon atom can be
inferred.
What are the different forms of EDTA?
EDTA is an amphiprotic substance, which means that it can both donate and accept protons. The four
hydrogen atoms shown in the above line structure are acidic, and because of this, the formula of EDTA is
often abbreviated H
4
Y, where H
4
represents the four acidic hydrogen atoms and Y represents the
remaining structure. The K
a
values for the sequential loss of these protons are 1, 0.032, 0.01, and
0.0022. The lone pairs of electrons on the nitrogen atoms in EDTA are capable of accepting protons.
The K
b
values for the protonation of these nitrogen atoms are 1.74 10
-4
and 1.45 10
-8
. EDTA can
therefore exist in many different forms depending on the pH of the solution. At very low pH, EDTA will
be present in its completely protonated form H
6
Y
2+
. At very high pH, EDTA will be present in its
completely deprotonated form, Y
4-
. At intermediate pH, EDTA will be present in one of its intermediate
forms. The following structures depict the various ionized forms of EDTA, along with their abbreviations
and the pH at which each predominates.



The negative log of an equilibrium constant is called a pK value. The pK
a
value, the negative log of the K
a

value, for an acid gives the pH at which there is an equal concentration of an acid and its conjugate base.
This can be seen by solving the Henderson-Hasselbalch equation for the pH at which an equal
concentration of an acid and its conjugate base would be present in a solution. The K
a
and K
b
values can
be used to determine the predominant form of an amphiprotic substance at different values of pH. The
following table was constructed using the pK
a
and pK
b
values for EDTA.
Form of
EDTA
H
6
Y
2+
H
5
H
+
H
4
HY H
3
Y
-
H
2
Y
2-
HY
3-
Y
4-


pH 0.0 1.5 2.0 2.66 6.16 10.24
Which form of EDTA do we start with?
EDTA is most frequently purchased as the dihdrate salt of its H
2
Y
2-
form, Na
2
H
2
Y2H
2
O (structure shown
below). Recall that hyrdrates are ionic compounds with loosely bound water molecules in their crystal
structure.


Which form of EDTA forms the most stable complex ions?
Very stable complex ions are formed between metal ions and EDTA in its completely deprotonated
form, Y
4-
. In order to get a significant portion of EDTA into this form, solutions of EDTA used in titrations
are typically buffered at high pH. In this experiment, you will use a buffer of NH
3
and NH
4
Cl to maintain
a pH of 10.

The completely deprotonated form of EDTA, Y
4-
, can form bonds through the lone pairs of electrons on
the 4 oxygen atom and 2 nitrogen atoms for a total of 6 binding sites. Because of the free rotation
about single (sigma) bonds EDTA is able to wrap around metal ions and orient these six binding sites
toward the metal ion. These six binding sites are arranged in the shape of an octahedron around the
metal ion as is shown in the above figure for the EDTA chelation of a metal ion.
How is EDTA used to determine the concentration of metal ions in a solution?
EDTA is able to form stable 1:1 complex ions with many different metal ions, and can therefore be used
in titrations to determine the concentrations of these metal ions in a solution. Because the reactions
used in these titrations involve the formation of complex ions, they are called complexiometric
titrations.
How do indicators in complexiometric titrations work?
Indicators used compexiometric titrations are complexing agents themselves. Unlike EDTA, however,
they are colored, and their color is different in their free and complexed forms. For example, the color
of Eriochrome black T, one of the indicators used in this experiment, is blue in its free form and wine red
when it is complexed with a metal ion. The color of these indicators is also affected by pH, which is
another reason that buffers are often used to regulate pH during complexiometric titrations. When the
indicator is added to water containing metal ions it will rapidly form a complex ion, and the solution will
take on the color of the complexed form of the indicator. As EDTA is added to the solution it will react
with the free metal ions in the solution first. After enough EDTA has been added to react with all of the
free metal ions, additional EDTA will displace the metal ions in the indicator-metal ion complex. This
occurs because the formation constant for the EDTA-metal ion complex is larger than that of the
indicator-metal ion complex. In other words, EDTA binds more strongly to the metal ions. As the
indicator loses its metal ions, the solution will become the color of the indicator in its free form. The
following summary and reactions illustrate this process.
Initially, indicator is added to a solution containing metal ions. The indicator reacts with the metal ions
in solution to form complex ions, and the solution takes on color of indicator-metal complex ion.

Prior to equivalence point, added EDTA reacts with free metal ions in solution to form complex ions.
The solution color does not change during this part of the titration.

As the equivalence point is approached, added EDTA displaces metal ions from the indicator-metal
complex ion, and the solution takes on color of the free indicator.

How can the individual concentrations of Ca
2+
and Mg
2+
be determined?
In this lab, you will use EDTA to determine the concentration of metal ions in a water sample. The most
common positively charged metal ions in natural waters are Ca
2+
and Mg
2+
. For this lab, you will assume
that these are the only metal ions present in your water. You will perform two sets of titrations. One
set of titrations will be used to determine the total concentration of Ca
2+
and Mg
2+
present in your water
sample. After selectively precipitating the Mg
2+
as Mg(OH)
2
, you will perform a second set of titrations
to determine the concentration of Ca
2+
in your water. The concentration of Mg
2+
in your water sample
can then be determined by difference.
Materials
EDTA: Na
2
H
2
EDTA2H
2
O (molar mass 372.24 g/mol), 0.6 g per student
pH 10 Buffer: Add 142 mL of 28 % (by mass) aqueous NH
3
to 17.5 g of NH
4
Cl and dilute to 250 mL with
deionized water.
Eriochrome black T indicator: Dissolve 0.2 g of the solid indicator in 15 mL of triethanolamine plus 5 mL
of absolute ethanol
50 % (by mass) aqueous NaOH
Water Standard: Evian bottled water or other brand if calcium and magnesium concentrations are
known.
Unknowns: Collect a water sample from a stream, lake, or ocean. Use a plastic bottle and fill it
completely to the top so that no air will be present in the sealed bottle. This will minimize the growth of
bacteria in your water.
Procedure
Solution preparation: If the EDTA has not been dried for you, start by drying about 1 g of
Na
2
H
2
EDTA2H
2
O (molar mass 372.24 g/mol) at 80C for 1 h and cool in a desiccator. Weigh out ~0.6 g
of the dry EDTA and record the mass to the nearest 0.0001 g. Add the EDTA to a 500 mL volumetric
flask and add about 400 mL of deionized water. Dissolve the EDTA with swirling and heating if
necessary. If you use heating, you will need to allow your EDTA to cool to room temperature before
diluting to the 500-mL mark on the neck of the volumetric flask. Use a squirt bottle of di H
2
O to careful
bring the volume of the solution to 500 mL.
Water standard: In order to test your reagents and the accuracy of the titration, you may first wish to
titrate a water sample with a known concentration of calcium and magnesium. Bottled mineral water
for which these concentrations are known can serve as a standard.
Unknown water samples: If you choose to analyze unknown water samples, you will need to determine
the appropriate sample size. The ability of titrations to precisely to determine unknown concentrations
relies on the ability to precisely deliver volumes with a buret. In order to deliver a volume to 4
significant digits using a buret, you must deliver at least 10 mL. Precision increases with the delivery of
higher volumes. Refilling the buret however will increase the number of volume readings that must be
made and therefore increase the uncertainty in determining the total volume delivered. The goal is to
deliver as much titrant as possible without having to refill the buret. Using 20-40 mL of titrant is a
reasonable goal. Because the concentrations of Mg
2+
and Ca
2+
in your unknown water sample are
unknown, it is impossible to know what sample volume will require 20-40 mL of titrant. Therefore, you
will need to perform some quick titrations (using the procedure below) with different sample volumes
to determine the appropriate sample size. Start with a 50-mL sample size and then adjust accordingly.
If the appropriate sample volume is less than 50 mL, bring the total volume up to ~50 mL by adding
sufficient deionized water. Once you have selected an appropriate sample size, perform careful
titrations on at least three samples of your unknown water.
Total Mg
2+
and Ca
2+
determination: Pipet a 50-mL sample of water into a 250-mL Erlenmyer flask. To
the sample add 3 mL of the pH 10 buffer and 6 drops of Eriochrome black T indicator. Rinse and fill a
clean 50-mL buret with your EDTA solution and record the initial volume. Titrate your water sample
until the color changes from wine red to blue. Perform a quick titration to determine the approximate
volume needed to reach the endpoint. In order to accurately identify the endpoint, where the last trace
of red has been removed from the solution, you may want to add small volumes of mineral water to
your titrated sample and practice reaching the endpoint. Once you have confidently identified the
endpoint, save this sample as a reference for subsequent titrations. Carefully titrate 3 more samples of
the water.
Blank titration: Titrate a 50-mL sample of the lab water that was used to prepare your solutions to
determine whether it contains any calcium or magnesium ions. If the lab water requires the addition of
EDTA to reach the endpoint, you will need to subtract this volume from each of your titration volumes.
Ca
2+
determination: If you wish to determine the individual concentration of calcium and magnesium in
your water sample, use the following procedures for precipitating the Mg
2+
and titrating the Ca
2+
that
remains in solution. Pipet four water samples into separate Erlenmyer flasks. To precipitate the Mg
2+
,
add 30 drops of 50% NaOH to each sample and swirl for 2 min. The precipitate may not be visible.
Because Eriochrome black T does not work well at this elevated pH, you will use hydroxynaphthol blue
as the indicator in these titrations. Add ~0.1 g of hydroxynaphthol blue to each sample. Perform one
quick titration to determine the approximate endpoint, and practice finding the endpoint if necessary.
To accurately determine the endpoint of each of the remaining samples, titrate to the blue endpoint and
then allow the sample to sit for 5 min with occasional swirling. This will allow any Ca(OH)
2
that may
have precipitated to redissolve. If the solution has turned back to red, add additional titrant to reach
the blue endpoint. Following the same procedure, perform a blank titration on a 50-mL sample of lab
water and make any volume corrections necessary.
Calculations: From the precise mass of EDTA that you weighed out, calculate the concentration of your
EDTA. Use the volume of EDTA needed to reach the endpoint in each of your titrations to determine
either the total concentration of Ca
2+
and Mg
2+

or the concentration of Ca
2+
alone. Recall that EDTA
reacts with either of these ions in a 1:1 mole ratio. If you performed the Ca
2+
titration, determine the
Mg
2+
concentration by difference from the total of Ca
2+
and Mg
2+
. Calculate the relative standard
deviation of replicate titrations. If bottled water was titrated, calculated the relative difference of your
Ca
2+
and Mg
2+
concentrations to those listed by the manufacturer.

Pre-lab Questions

Determination of Ca
2+
and Mg
2+

in Water by EDTA Titration

Name:_______________________
Instructor:____________________


1. If a student prepared an EDTA solution for a complexiometric titration by dissolving 0.5946 g of
Na
2
H
2
EDTA2H
2
O (molar mass 372.24 g/mol) in enough water to bring the total solution volume to 500.0
mL, what would the molar concentration of EDTA in the solution be? Clearly show any required
calculations with proper units and significant digits.





2. A student titrates a 50.00-mL sample of water with a 0.003125 M EDTA solution. If the titration
requires 34.64 mL of EDTA to reach the endpoint, what is the total concentration of Mg
2+
and Ca
2+
in the
water sample? Clearly show any required calculations with proper units and significant digits.





3. The same student then adds NaOH to another 50.00 mL sample of the same water and then titrates
again with EDTA. This time the titration requires 23.67 mL of EDTA solution. Determine the individual
concentrations of Ca
2+
and Mg
2+
in the water sample. Clearly show any required calculations with
proper units and significant digits.


Report Sheet

Determination of Ca
2+
and Mg
2+

in Water by EDTA Titration

Name:_______________________
Instructor:____________________

Mass of Na
2
H
2
EDTA2H
2
O used to prepare EDTA solution (g) _______________
Concentration of EDTA solution (M) ______________________
Water standard
Total Mg
2+
and Ca
2+
determination:
Volume of EDTA titrant
Initial Volume
(mL)
Final
Volume (mL)
Volume
Delivered (mL)
Volume for
blank (mL)
Corrected
Volume (mL)
Rough Trial

Trial 1
Trial 2
Trial 3

Blank

Total concentration of Ca
2+
and Mg
2+
(M)
Trial 1
Trial 2
Trial 3

Mean
Standard Deviation
Relative Standard Deviation (%)

Ca
2+
determination:
Volume of EDTA titrant
Initial Volume
(mL)
Final
Volume (mL)
Volume
Delivered (mL)
Volume for
blank (mL)
Corrected
Volume (mL)
Rough Trial

Trial 1
Trial 2
Trial 3

Blank

Concentration of Ca
2+
(M)
Trial 1
Trial 2
Trial 3

Mean
Standard Deviation
Relative Standard Deviation (%)

Total concentration of Ca
2+
and
Mg
2+
(M)
Mean Concentration of Ca
2+
(M) Concentration of Mg
2+
by
difference(M)


Comparison of manufacturer-reported and experimentally-determined Mg
2+
and Ca
2+
concentrations
Ca
2+
concentration (M) Mg
2+
concentration (M)
Reported by Manufacturer
Experimental
Relative Difference (%)

Sample calculations (use proper units and significant digits):
EDTA concentration:


Total concentration of Ca
2+
and Mg
2+
:


Concentration of Ca
2+
:


Concentration of Mg
2+
:
Report Sheet

Determination of Ca
2+
and Mg
2+

in Water by EDTA Titration

Name:_______________________
Instructor:____________________

Unknown water samples
Volume of water sample used for titration (mL)__________________________
Total Mg
2+
and Ca
2+
determination:
Volume of EDTA titrant
Initial Volume
(mL)
Final
Volume (mL)
Volume
Delivered (mL)
Volume for
blank (mL)
Corrected
Volume (mL)
Rough Trial

Trial 1
Trial 2
Trial 3

Blank

Total concentration of Ca
2+
and Mg
2+
(M)
Trial 1
Trial 2
Trial 3

Mean
Standard Deviation
Relative Standard Deviation (%)

Ca
2+
determination:
Volume of EDTA titrant
Initial Volume
(mL)
Final
Volume (mL)
Volume
Delivered (mL)
Volume for
blank (mL)
Corrected
Volume (mL)
Rough Trial

Trial 1
Trial 2
Trial 3

Blank


Concentration of Ca
2+
(M)
Trial 1
Trial 2
Trial 3

Mean
Standard Deviation
Relative Standard Deviation (%)

Total concentration of Ca
2+
and
Mg
2+
(M)
Mean Concentration of Ca
2+
(M) Concentration of Mg
2+
by
difference(M)


Sample calculations (use proper units and significant digits):
EDTA concentration:


Total concentration of Ca
2+
and Mg
2+
:


Concentration of Ca
2+
:


Concentration of Mg
2+
:

Post-lab questions


Determination of Ca
2+
and Mg
2+

in Water by EDTA Titration

Name:_______________________
Instructor:____________________

1. Write out the net ionic equation for the reaction that was used to remove Mg
2+
from your water
sample so that Ca
2+
alone could be determined.

Why is it that Mg
2+
is removed from solution but Ca
2+
is not? The K
sp
values for the hydroxides of Mg
2+

and Ca
2+
are 1.8 10
-11
and 6.5 10
-6
respectively.



2. Calcium Disodium Versenate (see structure below) is a form of EDTA that can be used to treat lead
poisoning in humans. In this drug, EDTA is present as a complex ion with Ca
2+
. How is it possible for this
drug to remove Pb
2+
from a persons blood if it is already complexed to Ca
2+
? The formation constants,
K
f
values, for the EDTA- Ca
2+
and EDTA- Pb
2+
complex ions are 4.9 10
10
and 1.1 10
18
respectively.





3. Two indicators, Funky Green (FG) and Infernal Orange (IO), are being considered for an EDTA titration
for the determination of Mg
2+
. The formation constants, K
f
values, for the indicator- Mg
2+
complex ion
are 2.34 10
5
for FG and 1.76 10
9
for IO. The formation constant for the EDTA-Mg
2+
complex ion is
6.17 10
8
. Which indicator would be the appropriate choice and why?


4. Challenge Question. Calcium Disodium Versenate (see structure below) is a form of EDTA that can
be used to treat lead poisoning in humans who have blood lead levels in the range of 20-70 micrograms
per deciliter. The recommended dosage of this drug for an adult is about 1000 mg. The average blood
volume of an adult is about 5 L. If the formation constant, K
f
, for the EDTA-Pb
2+
complex ion is 1.1
10
18
, what would be the concentration of free Pb
2+
in an average adults blood if she had an initial blood
Pb
2+
concentration of 50 micrograms per deciliter and received a 1000 mg dose of calcium disodium
versenate . Assume that there are no other competing reactions.