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
Note: multiple atoms from EDTA are binding Mn2+

Metal Lewis Acid or Electron-pair acceptor Ligand Lewis Base or Electron-pair donor

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 Elements that contain free electron pairs that may be donated to a metal
Fe-DTPA Complex

Most chelating agents contain N or O

-

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
channel contains pore that only allows K+ to pass K+ is chelated by O in channel

K+ channel spans membrane Opening of potassium channel allows K+ to exit cell and change the electrical potential across membrane

Current Opinion in Structural Biology 2001, 11:408414

http://www.bimcore.emory.edu/home/molmod/Wthiel/Kchannel.html

EDTA Titrations
Metal Chelate Complexes
1.) Formation Constant (Kf)

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 (Ksp), acid-base (Ka, Kb), water dissociation (Kw), etc

Chelate effect: ability of multidentate ligands to form stronger metal complexes compared to monodentate ligands.

Kf = 8x109

Kf = 4x109

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 Kf 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 Kf 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 (Y4-) is the one which primarily reacts with metal ions.

EDTA-Mn Complex

EDTA Titrations
EDTA
2.) Acid-Base Forms

Fraction ( ) of the most basic form of EDTA (Y4-) is defined by the H+ concentration and acid-base equilibrium constants
+

[ H

+[ H

K1+[ H

K1 K

+[ H

K3+[ H

K 1 K 2 K3 K K 2 K3 K4 K 5 K6 Y 4 = ]

+[ H

]K

+K

Fraction ( ) of EDTA in the form Y4-: 4

Y 4 =

[Y ] 2+ + 2 3 4 [ H 6 Y ]+[ H 5 Y ]+[ H 4 Y ]+[ H 3 Y ]+[ H 2 Y ]+[ HY ]+[ Y ]

[ Y 4 ] Y 4 = [ EDTA ]
where [EDTA] is the total concentration of all free EDTA species in solution

Y4- is depended on the pH of the solution

EDTA Titrations
EDTA
3.) EDTA Complexes

The basic form of EDTA (Y4-) reacts with most metal ions to form a 1:1 complex.
Other forms of EDTA will also chelate metal ions

[ MY n- 4 ] K f = n+ 4 [ M ][ Y ]
Note: This reaction only involves Y4-, but not the other forms of EDTA

Recall: the concentration of Y4- and the total concentration of EDTA is solution [EDTA] are related as follows: [ Y 4 ]= Y 4 EDTA

where Y4-is dependent on pH

EDTA Titrations
EDTA
3.) EDTA Complexes

The basic form of EDTA (Y4-) reacts with most metal ions to form a 1:1 complex.

EDTA Titrations
EDTA
3.) EDTA Complexes

Substitute [Y4-] into Kf equation

[Y

]= Y 4 [ EDTA ]

[ MY n- 4 ] K f = n+ 4 [ M ][ Y ]
where [EDTA] is the total concentration of EDTA added to the solution not bound to metal ions

[ MY n- 4 ] K f = n+ [ M ] Y 4-[ EDTA ]

If pH is fixed by a buffer, then Y4- is a constant that can be combined with Kf

Conditional or effective formation constant: (at a given pH)

[ MY ] ' K f = K = K f Y 4-= n+ [ M ][ EDTA ]

n- 4

EDTA Titrations
EDTA
3.) EDTA Complexes

Assumes the uncomplexed EDTA were all in one form

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

EDTA Titrations
EDTA
4.) Example:

What is the concentration of free Fe3+ 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
Kf decreases [Fe3+] = 5.4x10-7 at pH 2.0 -> [Fe3+] = 1.4x10-12 at pH 8.0

In order to get a complete titration (Kf 106), 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

Minimum pH for Effective Titration of Metal Ions

By adjusting the pH of an EDTA titration:

one type of metal ion (e.g. Fe3+) can be titrated without interference from others (e.g. Ca2+)

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)

K 'f = K f Y 4

The Titration Curve has three distinct regions:

Before the equivalence point (excess Mn+) At the equivalence point ([EDTA]=[Mn+] After the equivalence point (excess EDTA)

pM =log [ M n + ]

EDTA Titrations
EDTA Titration Curves
2.) Example

What is the value of [Mn+] and pM for 50.0 ml of a 0.0500 M Mg2+ 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?

Kf = 108.79 = 6.2x108 Y4- at pH 10.0 = 0.30

mL EDTA at equivalence point:

( V e ( mL ) ) ( 0 . 0500

M ) = ( 5 . 00 mL ) ( 0 . 0500 M ) V e = 50 . 00 mL

mmol of EDTA

mmol of Mg2+

EDTA Titrations
EDTA Titration Curves
2.) Example

(a) Before Equivalence Point ( 5.0 mL of EDTA)

Before the equivalence point, the [Mn+] is equal to the concentration of excess unreacted Mn+. Dissociation of MYn-4 is negligible.

moles of Mg2+ originally present

2+

moles of EDTA added

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

Original volume solution Volume titrant added
Dilution effect

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

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 MgY2-

Original volume of Original [Mn+] Mn+ solution

[ MgY 2 ]=( 0 . 0500 M ) ( 0 . 0500 L ) ( 0. 0500 L + 0. 0500 L )

Moles Mg+ moles MgY2-

Original volume solution

Volume titrant added

Dilution effect

[ MgY 2 ]=0.0250 M

EDTA Titrations
EDTA Titration Curves
2.) Example

(b) At Equivalence Point ( 50.0 mL of EDTA) The concentration of free Mg2+ is then calculated as follows:

Initial Concentration (M) Final Concentration (M)

0 x

0 x 2

0.0250 0.0250 - x

[ Mg ( EDTA ) ] ' K = K f Y 4 = f [ Mg 2+ ][ EDTA ]

( 6 . 2 10 8 )( 0 . 30 )=
Solve for x using the quadratic equation:

( 0 . 0250 x ) ( x )( x )

x=[ Mg 2 + ]=[ EDTA ]=1.16 10 5 pMg 2 + =4 .94

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 MgY2- and there is excess, unreacted EDTA. A small amount of free Mn+ exists in equilibrium with MgY4- and EDTA.

Calculate excess [EDTA]:

Volume excess Original [EDTA] titrant

[ EDTA ]= ( 0. 0500 M )( 0 .0010 L ) ( 0. 0500 L + 0. 0510 L )

Excess moles EDTA

Original volume solution

Volume titrant added

Dilution effect

[ EDTA ]= 4.95 104 M

EDTA Titrations
EDTA Titration Curves
2.) Example

(c) After the Equivalence Point ( 51.0 mL of EDTA)

Calculate [MgY2-]:

Original volume of Original [Mn+] Mn+ solution

[ MgY 2 ]( 0. 0500 M ) ( 0. 0500 L ) ( 0. 0500 L + 0 .0510 L )

Moles Mg+ moles MgY2Only Difference

Original volume solution

Volume titrant added

Dilution effect

[ MgY 2 ]0 .0248 M

EDTA Titrations
EDTA Titration Curves
2.) Example

(c) After the Equivalence Point ( 51.0 mL of EDTA)

[Mg2+-] is given by the equilibrium expression using [EDTA] and [MgY2-]:

[ Mg ( EDTA ) ] K ' = K f Y 4 = f [ Mg 2+ ][ EDTA ]

( 6 . 2 10 )( 0. 30 )=
8

( 0. 0248 M ) ( x )( 4 . 95 104 M )

x=[ Mg 2 + ]=2 .7 107 pMg 2 + = 6 .57

EDTA Titrations
EDTA Titration Curves
2.) Example

Final titration curve for 50.0 ml of 0.0500 M Mg2+ 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 Zn2+ vs. Mg2+. This is due to Zn2+ having a larger formation constant.

The completeness of these reactions is dependent on 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 Kf.
Larger change at the equivalence point.

Exception: If Mn+ reacts with OH- to form an insoluble metal hydroxide Auxiliary Complexing Agents: a ligand can be added that complexes with Mn+ strong enough to prevent hydroxide formation.
Ammonia, tartrate, citrate or triethanolamine Binds metal weaker than EDTA

Fraction of free metal ion ( M) depends on the equilibrium constants ( ) or cumulative formation constants:

M =

1 1 + 1 [ L ] + 2 [ L ]2 + n [ L ]n

Use a new conditional formation constant that incorporates the fraction of free metal:

K = Y 4 Zn2 + K f
f

''

EDTA Titrations
Auxiliary Complexing Agents
2.) Illustration:

Titration of Cu+2 (CuSO4) 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

CuSO4

Cu-ammonia Cu-EDTA

EDTA Titrations
Metal Ion Indicators
1.) Determination of EDTA Titration End Point

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

Potential Measurements

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

End Point indicated by a color change from red to blue

(red)

(colorless)

(colorless)

(blue)

EDTA Titrations
Metal Ion Indicators
2.) Illustration

Titration of Mg2+ by EDTA

Eriochrome Black T Indicator

Addition of EDTA

Before

Near Equivalence point

After

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) 2) 3) G. Schwarzenbach and H. Flaschka, Complexometric Titrations, Methuen:London, 1969. H.A. Flaschka, EDTA Titrations, Pergamon Press:New York, 1959 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) b) c) d) e) Direct Titrations Back Titrations Displacement Titrations Indirect Titrations 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. A known excess of EDTA is added to analyte
Free EDTA left over after all metal ion is bound with EDTA

3.) Back Titrations

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

f ( analyte ) Y 4 > K f ( sec ond metal ion )

Y 4

EDTA Titrations
EDTA Titration Techniques
4.) Displacement Titration

Used for some analytes that dont have satisfactory metal ion indicators Analyte (Mn+) is treated with excess Mg(EDTA)2-, causes release of Mg2+. Requires:

f ( M n+ )

Y 4 K

f ( Mg 2 + )

Y 4

Amount of Mg2+ released is then determined by titration with a standard EDTA solution Concentration of released Mg2+ equals [Mn+]

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. SO42- + excess Ba2+ 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 Mg2+ solution

[Total EDTA] = [MYn-4] + [Y4-]

complex free

Known

determine

Titrate

EDTA Titrations
EDTA Titration Techniques
6.) Masking Agents

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

Al3+ is not available to bind EDTA because of the complex with F -

Requires:

f ( AlF 3 6 )

>K

f ( Al ( EDTA ))

Demasking: refers to the release of a metal ion from a masking agent