EDTA Titrations

Introduction

1.) Metal Chelate Complexes

Any reagent which reacts with an analyte in a known ratio and with a large
equilibrium constant can potentially be used in a titration.

Complexation Titrations are based on the reaction of a metal ion with a
chemical agent to form a metal-ligand complex.
Metal
Ligand
Metal-Ligand Complex
Metal Lewis Acid or Electron-pair acceptor
Ligand Lewis Base or Electron-pair donor
Note: multiple atoms from
EDTA are binding Mn
2+
EDTA Titrations
Introduction

1.) Metal Chelate Complexes

Complexation Titrations are essentially a Lewis acid-base reaction, in which
an electron pair is donated from one chemical to another
The ligands used in complexometric titrations are also known as chelating
agents.
- Ligand that attaches to a metal ion through more than one ligand atom
Most chelating agents contain N or O
- Elements that contain free electron pairs that may be donated to a metal
Fe-DTPA Complex

EDTA Titrations
Metal Chelation in Nature

1.) Potassium Ion Channels in Cell Membranes

Electrical signals are essential for life
Electrical signals are highly controlled by the selective passage of ions across
cellular membranes
- Ion channels control this function
- Potassium ion channels are the largest and most diverse group
- Used in brain, heart and nervous system
Current Opinion in Structural Biology 2001, 11:408414
Opening of potassium channel allows K
+
to exit cell
and change the electrical potential across membrane
K
+
channel spans membrane
channel contains
pore that only
allows K
+
to pass
K
+
is chelated by O
in channel
http://www.bimcore.emory.edu/home/molmod/Wthiel/Kchannel.html
EDTA Titrations
Metal Chelate Complexes

1.) Formation Constant (K
f
)

The equilibrium constant for the reaction between a metal ion (M
+n
) and a
chelating agent (L
-P
) is known as a formation constant or stability constant.

Applying different and specific names to the general equilibrium constant is a
common occurrence
- Solubility (K
sp
), acid-base (K
a
, K
b
), water dissociation (K
w
), etc

Chelate effect: ability of multidentate ligands to form stronger metal
complexes compared to monodentate ligands.
K
f
= 8x10
9
K
f
= 4x10
9
2 ethylenediamine molecules binds tighter than 4 methylamine molecules
EDTA Titrations
Metal Chelate Complexes

2.) Chelate Effect

Usually chelating agents with more than one electron pair to donate will form
stronger complexes with metal ions than chelating agents with only one
electron pair.
- Typically more than one O or N
- Larger K
f
values

Multidentate ligand: a chelating agent with more than one free electron pair
- Stoichiometry is 1:1 regardless of the ion charge

Monodentate ligand: a chelating agent with only one pair of free electrons
Multidentate ligand that binds radioactive metal attached
to monoclonal antibody (mAb).

mAb is a protein that binds to a specific feature on a
tumor cell delivering toxic dose of radiation.
EDTA Titrations
EDTA

1.) EDTA (Ethylenediaminetetraacetic acid)

One of the most common chelating agents used for complexometric titrations
in analytical chemistry.

EDTA has 6 nitrogens & oxygens in its structure giving it 6 free electron pairs
that it can donate to metal ions.
- High K
f
values
- 6 acid-base sites in its structure
EDTA Titrations
EDTA

2.) Acid-Base Forms

EDTA exists in up to 7 different acid-base forms depending on the solution
pH.

The most basic form (Y
4-
) is the one which primarily reacts with metal ions.
EDTA-Mn Complex

EDTA Titrations
EDTA

2.) Acid-Base Forms

Fraction (o) of the most basic form of EDTA (Y
4-
) is defined by the H
+

concentration and acid-base equilibrium constants
| | EDTA
Y
Y HY Y H Y H Y H Y H Y H
Y
4
Y
4 3 2
2 3 4 5
2
6
4
Y
4
4
] [
] [ ] [ ] [ ] [ ] [ ] [ ] [
] [

+ +

=
+ + + + + +
=

o
o
Fraction (o) of EDTA in the form Y
4-
:
where [EDTA] is the total concentration of all free EDTA species in solution
} ] [ ] [ ] [ ] [ ] [ ] {[
2 3 4 5 6
6 5 4 3 2 1 5 4 3 2 1 4 3 2 1 3 2 1 2 1 1
6 5 4 3 2 1
Y
K K K K K K K K K K K H K K K K H K K K H K K H K H H
K K K K K K
4
+ + + + + +
=
+ + + + + +

o
o
Y4-
is depended on the pH of the solution
EDTA Titrations
EDTA

3.) EDTA Complexes

The basic form of EDTA (Y
4-
) reacts with most metal ions to form a 1:1
complex.
- Other forms of EDTA will also chelate metal ions








Recall: the concentration of Y
4-
and the total concentration of EDTA is
solution [EDTA] are related as follows:
] ][ [
] [
+
=
4 n
4 n-
f
Y M
MY
K
Note: This reaction only involves Y
4-
, but not the other forms of EDTA
| | EDTA Y
4
Y
4

=

o ] [
where o
Y4-
is dependent on pH
EDTA Titrations
EDTA

3.) EDTA Complexes

The basic form of EDTA (Y
4-
) reacts with most metal ions to form a 1:1
complex.
EDTA Titrations
EDTA

3.) EDTA Complexes

Substitute [Y
4-
] into K
f
equation










If pH is fixed by a buffer, then o
Y4-
is a constant that can be combined with K
f
] ][ [
] [
+
=
4 n
4 n-
f
Y M
MY
K
| | EDTA Y
4
Y
4

=

o ] [
] [ ] [
] [
- 4
Y
EDTA M
MY
K
n
4 n-
f
o
+
=
where [EDTA] is the total
concentration of EDTA added
to the solution not bound to
metal ions
] ][ [
] [
- 4
Y
EDTA M
MY
K K K
n
4 n-
f
'
f
+
= = = o
Conditional or effective formation constant:
(at a given pH)
EDTA Titrations
EDTA

3.) EDTA Complexes

Assumes the uncomplexed EDTA were all in one form

- 4
Y
o
f
'
f
K K =
at any pH, we can find o
Y4-
and evaluate K
f

EDTA Titrations
EDTA

4.) Example:

What is the concentration of free Fe
3+
in a solution of 0.10 M Fe(EDTA)
-
at pH
8.00?
EDTA Titrations
EDTA

5.) pH Limitation
Note that the metal EDTA complex becomes less stable as pH decreases
- K
f
decreases
- [Fe
3+
] = 5.4x10
-7
at pH 2.0 -> [Fe
3+
] = 1.4x10
-12
at pH 8.0

In order to get a complete titration (K
f
10
6
), EDTA requires a certain
minimum pH for the titration of each metal ion
End Point becomes less distinct as pH is
lowered, limiting the utility of EDTA as a titrant

EDTA Titrations
EDTA

5.) pH Limitation

By adjusting the pH of an EDTA
titration:
one type of metal ion (e.g. Fe
3+
) can
be titrated without interference from
others (e.g. Ca
2+
)
Minimum pH for Effective
Titration of Metal Ions

EDTA Titrations
EDTA Titration Curves

1.) Titration Curve

The titration of a metal ion with EDTA is similar to the titration of a strong acid
(M
+
) with a weak base (EDTA)




The Titration Curve has three distinct regions:
- Before the equivalence point (excess M
n+
)

- At the equivalence point ([EDTA]=[M
n+
]

- After the equivalence point (excess EDTA)
- 4
Y
o
f
'
f
K K =
] [
+
=
n
M log pM
EDTA Titrations
EDTA Titration Curves

2.) Example

What is the value of [M
n+
] and pM for 50.0 ml of a 0.0500 M Mg
2+
solution
buffered at pH 10.00 and titrated with 0.0500 m EDTA when (a) 5.0 mL, (b)
50.0 mL and (c) 51.0 mL EDTA is added?
K
f
= 10
8.79
= 6.2x10
8
o
Y4-
at pH 10.0 = 0.30

( )( ) ( ) mL 00 . 50 V ) M 0500 . 0 ( mL 00 . 5 M 0500 . 0 ) mL ( V
e e
= =
mL EDTA at equivalence point:

mmol of EDTA
mmol of Mg
2+
EDTA Titrations
EDTA Titration Curves

2.) Example

(a) Before Equivalence Point ( 5.0 mL of EDTA)
Before the equivalence point, the [M
n+
] is equal to the concentration of excess
unreacted M
n+
. Dissociation of MY
n-4
is negligible.

] [
)] )( ( - ) )( [(
] [
L 0050 . 0 L 0500 . 0
L 0050 . 0 M EDTA 0500 . 0 L 0500 . 0 M Mg 0500 . 0
Mg
2
2
+
=
+
+
moles of Mg
2+

originally present
moles of EDTA added

Original volume
solution

Volume titrant
added

39 . 1 Mg log pMg M 0409 . 0 Mg
2 2 2
= = =
+ + +
] [ ] [
Dilution effect

EDTA Titrations
EDTA Titration Curves

2.) Example

(b) At Equivalence Point ( 50.0 mL of EDTA)
Virtually all of the metal ion is now in the form MgY
2-
) (
) (
) ( ] [
L 0500 . 0 L 0500 . 0
L 0500 . 0
M 0500 . 0 MgY
2
+
=

Original [M
n+
]

Original volume of
M
n+
solution

Original volume
solution

Volume titrant
added

Dilution effect

Moles Mg
+
moles MgY
2-
M 0250 . 0 MgY
2
=

] [
EDTA Titrations
EDTA Titration Curves

2.) Example

(b) At Equivalence Point ( 50.0 mL of EDTA)
The concentration of free Mg
2+
is then calculated as follows:

Initial Concentration (M) 0 0 0.0250
Final Concentration (M) x x 0.0250 - x
] ][ [
] ) [
EDTA Mg
EDTA ( Mg
K K
2
2 -
Y
f
'
4
f +
= =

o
) x )( x (
) x 0250 . 0 (
) 30 . 0 )( 10 2 . 6 (
8

=
Solve for x using the quadratic equation:

94 . 4 pMg 10 16 . 1 EDTA Mg x
2 5 2
= = = =
+ +
] [ ] [
EDTA Titrations
EDTA Titration Curves

2.) Example

(c) After the Equivalence Point ( 51.0 mL of EDTA)
Virtually all of the metal ion is now in the form MgY
2-
and there is excess,
unreacted EDTA. A small amount of free M
n+
exists in equilibrium with
MgY
4-
and EDTA.
) (
) )( (
] [
L 0510 . 0 L 0500 . 0
L 0010 . 0 M 0500 . 0
EDTA
+
=
Original [EDTA]

Volume excess
titrant

Original volume
solution

Volume titrant
added

Dilution effect

Excess moles EDTA

M 10 95 . 4 EDTA
4
= ] [
Calculate excess [EDTA]:
EDTA Titrations
EDTA Titration Curves

2.) Example

(c) After the Equivalence Point ( 51.0 mL of EDTA)
Calculate [MgY
2-
]:
) (
) (
) ( ] [
L 0510 . 0 L 0500 . 0
L 0500 . 0
M 0500 . 0 MgY
2
+
~

Original [M
n+
]

Original volume of
M
n+
solution

Original volume
solution

Volume titrant
added

Dilution effect

Moles Mg
+
moles MgY
2-
M 0248 . 0 MgY
2
~

] [
Only Difference

EDTA Titrations
EDTA Titration Curves

2.) Example

(c) After the Equivalence Point ( 51.0 mL of EDTA)
[Mg
2+-
] is given by the equilibrium expression using [EDTA] and [MgY
2-
]:
] ][ [
] ) [
EDTA Mg
EDTA ( Mg
K K
2
2 -
Y
f
'
4
f +
= =

o
) M 10 95 . 4 )( x (
) M 0248 . 0 (
) 30 . 0 )( 10 2 . 6 (
4
8

=
57 . 6 pMg 10 7 . 2 Mg x
2 7 2
= = =
+ +
] [
EDTA Titrations
EDTA Titration Curves

2.) Example

Final titration curve for 50.0 ml of 0.0500 M Mg
2+
with 0.0500 m EDTA at pH
10.00.
- Also shown is the titration of 50.0 mL of 0.0500 M Zn
2+
Note: the equivalence point is sharper for Zn
2+

vs. Mg
2+
. This is due to Zn
2+
having a larger
formation constant.
The completeness of these reactions is
dependent on o
Y4-
and correspondingly pH.
pH is an important factor in setting the completeness
and selectivity of an EDTA titration
EDTA Titrations
Auxiliary Complexing Agents

1.) Metal Hydroxide

In general, as pH increases a titration of a metal ion with EDTA will have a
higher K
f
.
- Larger change at the equivalence point.

Exception: If M
n+
reacts with OH
-
to form an insoluble metal hydroxide

Auxiliary Complexing Agents: a ligand can be added that complexes with M
n+

strong enough to prevent hydroxide formation.
- Ammonia, tartrate, citrate or triethanolamine
- Binds metal weaker than EDTA
f
Zn Y
' '
K K
2 4
f
+
= o o
n
n
2
2 1
M
] L [ ] L [ ] L [ 1
1
| | |
o
+ + +
=

Fraction of free metal ion (o

M
) depends on the
equilibrium constants (|) or cumulative formation
constants:
Use a new conditional formation constant that
incorporates the fraction of free metal:
EDTA Titrations
Auxiliary Complexing Agents

2.) Illustration:

Titration of Cu
+2
(CuSO
4
) with EDTA

Addition of Ammonia Buffer results in a dark blue solution
- Cu(II)-ammonia complex is formed
Addition of EDTA displaces ammonia with corresponding color change
CuSO
4
Cu-EDTA

Cu-ammonia

EDTA Titrations
Metal Ion Indicators

1.) Determination of EDTA Titration End Point

Four Methods:
1. Metal ion indicator
2. Mercury electrode
3. pH electrode
4. Ion-selective electrode

Metal Ion Indicator: a compound that changes color when it binds to a metal
ion
- Similar to pH indicator, which changes color with pH or as the compound
binds H
+

For an EDTA titration, the indicator must bind the metal ion less strongly than
EDTA
- Similar in concept to Auxiliary Complexing Agents
- Needs to release metal ion to EDTA
Potential
Measurements
(red) (colorless) (colorless)
(blue)
End Point indicated by a color
change from red to blue
EDTA Titrations
Metal Ion Indicators

2.) Illustration

Titration of Mg
2+
by EDTA
- Eriochrome Black T Indicator

Addition of EDTA
Before Near After
Equivalence point
EDTA Titrations
Metal Ion Indicators

3.) Common Metal Ion Indicators

Most are pH indicators and can only be used over a given pH range
EDTA Titrations
Metal Ion Indicators

3.) Common Metal Ion Indicators

Useful pH ranges
EDTA Titrations
EDTA Titration Techniques

1.) Almost all elements can be determined by EDTA titration

Needs to be present at sufficient concentrations

Extensive Literature where techniques are listed in:
1) G. Schwarzenbach and H. Flaschka, Complexometric Titrations,
Methuen:London, 1969.
2) H.A. Flaschka, EDTA Titrations, Pergamon Press:New York, 1959
3) C.N. Reilley, A.J. Bernard, Jr., and R. Puschel, In: L. Meites (ed.) Handbook of
Analytical Chemistry, McGraw-Hill:New York, 1963; pp. 3-76 to 3-234.

Some Common Techniques used in these titrations include:
a) Direct Titrations
b) Back Titrations
c) Displacement Titrations
d) Indirect Titrations
e) Masking Agents
EDTA Titrations
EDTA Titration Techniques

2.) Direct Titrations

Analyte is buffered to appropriate pH and is titrated directly with EDTA

An auxiliary complexing agent may be required to prevent precipitation of
metal hydroxide.

3.) Back Titrations
A known excess of EDTA is added to analyte
- Free EDTA left over after all metal ion is bound with EDTA

The remaining excess of EDTA is then titrated with a standard solution of a
second metal ion

Approach necessary if analyte:
- precipitates in the presence of EDTA
- Reacts slowly with EDTA
- Blocks the indicator

Second metal ion must not displace analyte from EDTA

>
4 4
Y
) ion metal ond (sec f
Y
) analyte ( f
K K o o
EDTA Titrations
EDTA Titration Techniques

4.) Displacement Titration

Used for some analytes that dont have satisfactory metal ion indicators

Analyte (M
n+
) is treated with excess Mg(EDTA)
2-
, causes release of Mg
2+
.









Amount of Mg
2+
released is then determined by titration with a standard EDTA
solution
- Concentration of released Mg
2+
equals [M
n+
]

+ +
>
4 2 4 n
Y ) Mg ( f Y ) M ( f
K K o o
Requires:
EDTA Titrations
EDTA Titration Techniques

5.) Indirect Titration

Used to determine anions that precipitate with metal ions

Anion is precipitated from solution by addition of excess metal ion
- ex. SO
4
2-
+ excess Ba
2+
- Precipitate is filtered & washed

Precipitate is then reacted with excess EDTA to bring the metal ion back into
solution

The excess EDTA is titrated with Mg
2+
solution

[Total EDTA] = [MY
n-4
] + [Y
4-
]
complex free
Known Titrate
determine
EDTA Titrations
EDTA Titration Techniques

6.) Masking Agents

A reagent added to prevent reaction of some metal ion with EDTA









Demasking: refers to the release of a metal ion from a masking agent
Al
3+
is not available to bind EDTA because of the complex with F
-
)) EDTA ( Al ( f
) AlF ( f
K K
3
6
>

Requires: