Acid-Base Titrations: Experimental Measurements of PK

Acid-Base Titrations
Introduction
1.) Experimental Measurements of pKa

pKa of amino acids in an active-site of a protein are related to its function
- Protein structure and environment significantly perturb pKa values
In medicinal chemistry, pKa and lipophilicity of a candidate drug predict how

easily it will cross a cell membrane
- Higher charge harder to cross membrane not a good drug
Introduction
2.) Example:
impact of the Asp on the pKa of His in the His-Asp catalytic dyad.
- Glucose 6-phosphate dehydrogenase (G6PD) catalyzes the oxidation of
glucose 6-phosphate using NAD+ or NADP+
- His-240 is the general base that extracts a proton from the C1-OH of G6P
The pKa of His-240 in the G6PD apoenzyme is found to be 6.4, which corresponds to an
unidentified pKa value of 6.3 that was previously derived from the dependence of kcat on pH.
These results suggest that the pKa of His-240 is unperturbed by Asp.
Biochemistry, Vol. 41, No. 22, 2002 6945

Introduction
3.) Overview
Titrations are Important tools in providing quantitative and qualitative data for
a sample.
To best understand titrations and the information they provide, it is necessary

to understand what gives rise to the shape of a typical titration curve.
To do this, acid-base equilibria are used to predict titration curve shapes.
proton release from PAA decreases with

increase in the degree of dissociation
for the highest polymer concentration
conformational change of the PAA from rod-

like conformation to a random coil form,
J. Phys. Org. Chem. 2006; 19: 129135

Titration of Strong Base with Strong Acid
1.) Graph of How pH changes as Titrant is Added

Assume strong acid and base completely dissociate
Any amount of H+ added will consume a

stoichiometric amount of OH-
K 1 K 10 14
w
Reaction Assumed to go to completion
Three regions of the titration curve

- Before the equivalence point, the pH is determined
by excess OH- in the solution
- At the equivalence point, H+ is just sufficient to

react with all OH- to make H2O
- After the equivalence point, pH is determined by

excess H+ in the solution.
1.) Graph of How pH changes as Titrant is

Added
Remember, equivalence point is the
ideal goal
Actually measure End Point

- Marked by a sudden physical
change: color, potential
Different Regions require different kinds

of calculations
- Illustrated examples
The true titration reaction is:
Titrant Analyte
2.) Volume Needed to Reach the Equivalence Point

Titration curve for 50.00 mL of 0.02000 M KOH with 0.1000 M HBr
At equivalence point, amount of H+ added will equal initial amount of OH-
Ve ( mL ) 0.1000 M 50.00 mL ( 0.02000 M ) Ve 10.00 mL

mmol of HBr mmol of OH-
at equivalence point being titrated
When 10.00 mL of HBr has been added, the titration is complete.

Prior to this point, there is excess OH- present.
After this point there is excess H+ present.
3.) Before the Equivalence Point

- Equivalence point (Ve) when 10.00 mL of HBr has been added
- When 3.00 mL of HBr has been added, reaction is 3/10 complete
Initial volume of OH-

Calculate Remaining [OH-]:
10.00 3.00 50.00

[OH ] 0.02000 M 0.0132 M
10.00 50.00 3.00
Fraction of OH
Initial
-
concentration
Dilution Factor Total volume
Remaining of OH -
Calculate [H+] and pH:
Kw 1.0 10 14 13
[H ] -
7 . 58 10 M pH 12.12
[OH ] 0.0132
4.) At the Equivalence Point

- Just enough H+ has been added to consume OH-
- pH determined by dissociation of water
Kw
Kw= 1x10-14
x x
Kw x 2 x 1.00 10 7 M pH 7.00
- pH at the equivalence point for any strong acid with strong base is 7.00
- Not true for weak acid-base titration
5.) After the Equivalence Point

- Adding excess HBr solution
- When 10.50 mL of HBr is added
Calculate volume of excess H+:

Vadded Vequivalence 10.50 10.00 0.50 mL
Calculate excess [H+]:

Volume of excess H+
0.50
[H ] 0.1000 M 4
8.26 10 M
50.00 10.50
Initial
concentration Dilution factor Total volume
Calculate pH: of H+
pH -log[H ] log( 8.26 10 4 M ) 3.08

6.) Titration Curve

Rapid Change in pH Near Equivalence Point
- Equivalence point is where slope is greatest
- Second derivative is 0
pH at equivalence point is 7.00, only for strong

acid-base
- Not True if a weak base-acid is used
Titration of Weak Acid with Strong Base
1.) Four Regions to Titration Curve

Before any added base, just weak acid (HA) in water
- pH determined by Ka
Ka
With addition of strong base buffer

- [ A ] equation
pH pK a log
pH determined by Henderson Hasselbach
[HA]

At equivalence point, all HA is converted into A-

- Kb
Weak base with pH determined by Kb
1.) Four Regions to Titration Curve

Beyond equivalence point, excess strong base is added to A- solution
- pH is determined by strong base
- Similar to titration of strong acid with strong base
2.) Illustrated Example:

Titration of 50.00 mL of 0.02000 M MES with 0.1000 M NaOH
- MES is a weak acid with pKa = 6.27
- Reaction goes to completion with addition of strong base

7
K 1K 1K / K 1 5 . 4 10
b w a 1 10 14 / 10 6.27
3.) Volume Needed to Reach the Equivalence Point

- Reaction goes to completion with addition of strong base
- Strong plus weak react completely
Ve ( mL ) 0.1000 M 50.00 mL ( 0.02000 M ) Ve 10.00 mL

mmol of base mmol of HA
4.) Region 1: Before Base is Added

Simply a weak-acid problem
Ka
Calculate [H+]: Ka= 10-6.27
F-x x x
x2 x2
K a x [H ] 1.03 10 4
F x 0.02000 x
Calculate pH:
pH - log[H ] log( 1.03 10 4 M ) 3.99

5.) Region 2: Before the Equivalence Point

Adding OH- creates a mixture of HA and A- Buffer
Calculate pH from [A-]/[HA] using Henderson-Hasselbach equation
Simply the difference
Calculate [A-]/HA]: Simply
of initialratio of volumes
quantities
Amount of added NaOH is 3 mL with equivalence point is 10 mL
Relative Initial quantities (HA1) 1 3 - -

10
7 3 3
Relative Final quantities - 10
10 10
Calculate pH:
[ A ] 3
pH pK a log 6.27 log 10 5.90
[HA] 7
10
5.) Region 2: Before the Equivalence Point

pH = pKa when the volume of titrant equals Ve

2
1 1 1
Relative Final quantities -
2 2 2
[ A ] 1
pH pK a log pK log 2 pK
[HA] a 1 a
2
5.) Region 3: At the Equivalence Point

Exactly enough NaOH to consume HA
Relative Final quantities - - 1 1
The solution only contains A- weak base

Kb Kw
Kb
Ka
F-x x x
x2 K
Kb w
Fx Ka

Calculate Formal concentration of [A-]:

A- is no longer 0.02000 M, diluted by the addition of NaOH
Initial volume of HA
50.00
F' 0.02000 M 0.0167 M
50 . 00 10 . 00
Initial
of HA

Calculate [OH-]:
x2 Kw 1 10 14
Kb 1.86 10 8
Fx Ka 10 6.27
x2
1.86 10 8 x [OH - ] 1.76 10 -5 M
0.0167 x
Calculate pH:
Kw 1 10 14

pH - log[H ] log log 9.25
x 0.0167

pH at equivalence
point is not 7.00
pH will always be above 7.00 for titration of a weak acid
because acid is converted into conjugate base at the equivalence point
5.) Region 4: After the Equivalence Point

Adding NaOH to a solution of A-
- NaOH is a much stronger base than A-
- pH determined by excess of OH-
Calculate volume of excess OH-:
Amount of added NaOH is 10.10 mL with equivalence point is 10 mL
Volume of excess OH-

Calculate excess [OH ]:-
0.10
[OH - ] 0.1000 M 1.66 10 4 M
50.00 10.10
Initial Total volume
concentration Dilution factor
Calculate pH: of OH-
Kw 1.00 10 14
pH - log[H ] log log 10.22
[OH ] 1.66 10 4

5.) Titration Curve

Two Important Features of the Titration Curve
Equivalence point: [OH-] = [HA]

Steepest part of curve
Maximum slope
pH=pKa
Maximum Buffer
Vb = Ve
Capacity
Minimum slope
5.) Titration Curve

Depends on pKa or acid strength
Inflection point or maximum slope decreases with weaker acid
- Equivalence point becomes more difficult to identify
weak acid small slope change in

titration curve
Difficult to detect equivalence point

Strong acid large slope change in
titration curve
Easy to detect equivalence point

5.) Titration Curve

Depends on acid concentration
Inflection point or maximum slope decreases with
lower acid concentration
- Equivalence point becomes more difficult to
identify
- Eventually can not titrate acid at very low
concentrations
High
Lowconcentration
concentrationlarge
smallslope
slopechange
changeinin
titration
titrationcurve
curve
Easy to detect
Difficult equivalence
to detect point
equivalence point
At low enough concentration, can not detect change
Titration of Weak Base with Strong Acid
1.) Simply the Reverse of the Titration of a Weak Acid with a Strong Acid
Again, Titration Reaction Goes to Completion:
Again, Four Distinct Regions to Titration Curve
Before acid is added just weak base reaction

- pH determined from Kb
Kb
F-x x x
[ A ]
Before equivalence pK
pHpoint, log
a buffer
- [HA] equation
pH determined from Henderson Hasselbach

+
Titration in Diprotic Systems
1.) Principals for Monoprotic Systems Apply to Diprotic Systems

Multiple equivalence points and buffer regions
Multiple Inflection Points in Titration Curve
Two equivalence points Kb1
Kb2
2.) A Typical Case

Titration of 10.0 mL of 0.100 M base (B) with 0.100 M HCl
- pKb1 = 4.00 and pKb2 = 9.00
Volume at First Equivalence Point (Ve)
Ve ( mL ) 0.100 M 10.00 mL ( 0.1000M ) Ve 10.00 mL

mmol of HCl mmol of B
Volume at Second Equivalence Point Must Be 2Ve

- Second reaction requires the same number of moles of HCl
2.) A Typical Case

Point A
- Before Acid Added
- Weak base problem
Kb1
0.100 - x x x
x2
1.00 10 4 x 3.11 10 3
0.100 x
Kw
[H ] pH 11 .49
x
2.) A Typical Case

Point between A &B
- Before First Equivalence Point
- Buffer problem
Kw 1 10 14 10
K a2 1 10
K b1 1 10 4
Point (1.5 mL) is before first

equivalence point (10 mL)
[ B] 10.00 ml 1.5 ml 8.5

5.67
[ BH ] 1.5 ml 1.5
[ B]
pH pK a 2 log 10.00 log 5.67 10.75
[ BH ]

2.) A Typical Case

Point B
- Before First Equivalence Point
- Buffer problem
Kw 1 10 14 10
K a2 1 10
K b1 1 10 4
Point B (5 mL) is halfway to first

equivalence point (10 mL)
[B] 10.00 ml 5 ml 5

1
[BH ] 5 ml 5
[B]
pH pK a 2 log 10.00 log 1 10.00
[BH ]

pH = pKa2=10.00
2.) A Typical Case

Point C
- First Equivalence Point
- Intermediate form of the Diprotic
acid
K 2 K1F K1Kw
[H ]
K1 F
Account for dilution for formal

Solve for pH using
concentration (F) of BH+
intermediate form equation Initial volume of B
10.00
F' 0(.10
1000
5 M
)( 10
10 )( 0.0500 ) ( 10 )( 10
5 14 0.0500 M
[H ] 105
.00 10.00 ) 3.16 x10 8
10 0.0500
Initial
pH log( 3.16 x10 8 ) 7.50
of B
2.) A Typical Case

Point D
- Before Second Equivalence
Point
- BufferProblem
14
Kw 1 10 5
K a2 1 10
K b1 1 10 9
Point D (15 mL) is halfway to second

equivalence point (2x10 mL). First,
subtract Ve (10 mL)
[BH ] 10.00 ml 5.00 ml 5
2
1
[BH2 ] 5 ml 5
[BH ]
pH pK a 2 log 5.00 log 1 5.00
[BH ]
2
2
pH = pKa1=5.00
2.) A Typical Case

Point E
- Second Equivalence Point
- Weak acid problem
Account for dilution for formal

concentration (F) of BH2+2 Initial volume of B
10.00
F' 0.1000 M 0.0333 M
20 . 00 10 . 00
Initial
of B
pH determined by acid dissociation of BH2+2
Kb2
2.) A Typical Case

Point E
- Second Equivalence Point
- Weak acid problem
Ka1 Kw
K a1
K b2
0.0333 - x x x
x2 x2
1.0 10 5
F x 0.0333 x
x 5.72 10 4 pH 3.24
2.) A Typical Case

Beyond Point E
- Past Second Equivalence Point
- Strong acid problem
pH from volume of strong acid added.

Addition of 25.00 mL:
Excess acid:
Concentration of H+:
5.00
[H ] 0.1000 M 2
1.43 10 M
25.00 10.00
pH:
pH - log [H ] log( 1.43 10 2 ) 1.85

3.) Blurred End Points

Two or More Distinct Equivalence
Points May Not be Observed in
Practice
- Depends on relative difference in
Kas or Kbs
- Depends on Relative strength of

Kas or Kbs
Only one Equivalence point is clearly evident
Second Ka is too strong and is not

a weak acid relative to titrant
End
End point:
point: 1st 2derivative
nd
derivative is zero
is maximum
4.) Using Derivatives to Find End Point
Useful when End points overlap
End Point of titration curve is where slope is
greatest
- dpH/dV is large
- pH change in pH between consecutive points
- V average of pair of volumes
- Second derivative is similar difference using
ph = 4.400-4.245=0.155 first derivative values
5.) Using Gran Plot to Find End Point

Method of Plotting Titration Data to Give a Linear
Relationship
A graph of Vb10-pH versus Vb is called a Gran plot

Vb 10 pH HA K a Ve Vb
A
where: Vb = volume of strong base added
Ve = volume of base needed to reach equivalence point
A-, HA = activity coefficients 1
5.) Using Gran Plot to Find End Point

Plot is a straight line
- If ratio of activity coefficients is constant
- Slope = -Ka HA/ a-
- X-intercept = Ve (must be extrapolated)
Measure End Point with data Before Reach End Point
Only use linear region of Gran Plot
- Changing ionic strength changes activity coefficients
- added salt to maintain constant ionic strength
Slope Gives Ka
x-intercept gives Ve
Never Goes to Zero, approximation that every

mole of OH- generates one mole of A- is not
true as Vb approaches Ve
End Point Determination
1.) Indicators: compound added in an acid-base titration to allow end

point detection
Common indicators are weak acids or bases
Different protonated species have different colors
1.) Indicators: compound added in an acid-base titration to allow end

point detection
Color Change of Thymol Blue between pH 1 and 11
pK = 1.7 pK = 8.9
2.) Choosing an Indicator

Want Indicator that changes color in the vicinity of the equivalence point
and corresponding pH
The closer the two match, the more accurate determining the end point will
be
Bromocresol purple color change

brackets the equivalence point and
is a good indicator choice
Bromocresol green will change color

Significantly past the equivalence
point resulting in an error.
2.) Choosing an Indicator
The difference between the end point

(point of detected color change) and the
true equivalence point is the indicator
error
Amount of indicator added should be

negligible
Indicators cover a range of pHs

3.) Example:
a) What is the pH at the equivalence point when 0.100 M hydroxyacetic acid

is titrated with 0.0500 M KOH?
b) What indicator would be a good choice to monitor the endpoint?

Acid-Base Titrations: Experimental Measurements of PK

Transféré par

Informations du document

Titre original

Copyright

Formats disponibles

Partager ce document

Partager ou intégrer le document

Options de partage

Avez-vous trouvé ce document utile ?

Ce contenu est-il inapproprié ?

Droits d'auteur :

Formats disponibles

Acid-Base Titrations: Experimental Measurements of PK

Transféré par

Droits d'auteur :

Formats disponibles

Acid-Base Titrations

1.) Experimental Measurements of pKa

In medicinal chemistry, pKa and lipophilicity of a candidate drug predict how

Biochemistry, Vol. 41, No. 22, 2002 6945

To best understand titrations and the information they provide, it is necessary

To do this, acid-base equilibria are used to predict titration curve shapes.

proton release from PAA decreases with

conformational change of the PAA from rod-

J. Phys. Org. Chem. 2006; 19: 129135

1.) Graph of How pH changes as Titrant is Added

Any amount of H+ added will consume a

Reaction Assumed to go to completion

Three regions of the titration curve

- At the equivalence point, H+ is just sufficient to

- After the equivalence point, pH is determined by

1.) Graph of How pH changes as Titrant is

Actually measure End Point

Different Regions require different kinds

The true titration reaction is:

2.) Volume Needed to Reach the Equivalence Point

At equivalence point, amount of H+ added will equal initial amount of OH-

Ve ( mL ) 0.1000 M 50.00 mL ( 0.02000 M ) Ve 10.00 mL

When 10.00 mL of HBr has been added, the titration is complete.

3.) Before the Equivalence Point

Initial volume of OH-

10.00 3.00 50.00

Calculate [H+] and pH:

4.) At the Equivalence Point

5.) After the Equivalence Point

Calculate volume of excess H+:

Calculate excess [H+]:

pH -log[H ] log( 8.26 10 4 M ) 3.08

6.) Titration Curve

pH at equivalence point is 7.00, only for strong

1.) Four Regions to Titration Curve

With addition of strong base buffer

At equivalence point, all HA is converted into A-

1.) Four Regions to Titration Curve

2.) Illustrated Example:

- Reaction goes to completion with addition of strong base

3.) Volume Needed to Reach the Equivalence Point

Ve ( mL ) 0.1000 M 50.00 mL ( 0.02000 M ) Ve 10.00 mL

4.) Region 1: Before Base is Added

pH - log[H ] log( 1.03 10 4 M ) 3.99

5.) Region 2: Before the Equivalence Point

Relative Initial quantities (HA1) 1 3 - -

5.) Region 2: Before the Equivalence Point

Relative Initial quantities (HA1) 1 1 - -

5.) Region 3: At the Equivalence Point

Relative Initial quantities (HA1) 1 1 - -

Relative Final quantities - - 1 1

The solution only contains A- weak base

5.) Region 3: At the Equivalence Point

Calculate Formal concentration of [A-]:

5.) Region 3: At the Equivalence Point

5.) Region 4: After the Equivalence Point

Volume of excess OH-

5.) Titration Curve

Equivalence point: [OH-] = [HA]

5.) Titration Curve

weak acid small slope change in

Difficult to detect equivalence point

Easy to detect equivalence point