pH and buffer

pH is a measure of the acidity or basicity of a solution.

Danish chemist Sren Peder Lauritz Srensen in 1909 :
Concept of p[H] and revised to the modern pH in 1924.
A.O.Beckman(1036) : Radiometer in Denmark
1070: Portable digital pHmeter
Electromotive force in cells is depended on activity rather than
concentration of hydrogen ions
pH - Negative decimal logarithm of the hydrogen ion activity in a
solution.
Ah - activity of hydrogen ions in units of Mol/L (molar
concentration).

Activity - sense of concentration,
activity is always less than the concentration ,defined as a
concentration (Mol/L) of an ion multiplied by activity coefficient.
Measurement of pH - by means of a glass electrode connected to a
milli-voltmeter
Measures the potential difference, or electromotive force,E, between
an electrode sensitive to the hydrogen ion activity and a reference
electrode, such as a calomel electrode or a silver chloride electrode.
Glass electrode is combined with the reference electrode and a
temperature sensor in one body.
The glass electrode relatively good (95 - 99.9%) follows the Nernst
equation:




E - measured potential , E0 - standard electrode potential, the
electrode potential for the standard state in which the activity is
one. R - gas constant, T - temperature in kelvins, F - Faraday
constant and n - number of electrons transferred (ion charge)
The electrode potential, E directly proportion to logarithm of the
hydrogen ion activity.
E = 0.059 log 1/ ( H+)
= 0.059 (- log H+)
= 0.059 pH
pH= 0.059 / E
0.06 V / pH unit change in voltmeter
Calomel electrode : pH=E 0.246 /0.059
The reduction reaction occurring at the calomel electrode
corresponds to the reduction of mercury (I).


The standard electrode potential for the test solution is then given by:




pH indicator

pH indicators - weak acids or weak bases.
HInd + H2O H3O+ + Ind-
HInd - the acid form
Ind- the conjugate base of the indicator.
It is the ratio of these that determines the color of the solution and
that connects the color to the pH value. For pH indicators that are
weak protolytes, the Henderson-Hasselbalch equation :



pOH
pOH - as a measure of the concentration of hydroxide ions, OH,
or alkalinity.
pOH is not measured independently, but is derived from pH. The
concentration of hydroxide ions in water is related to the
concentration of hydrogen ions by
[OH] = KW /[H+]
where KW is the self-ionisation constant of water.
Taking logarithms
[OH] = KW /[H+]

pOH = pKW pH.
At room temperature pOH 14 pH.
Buffer - an aqueous solution consisting of a
mixture of a weak acid and its conjugate base or
a weak base and its conjugate acid.
pH of the solution changes very little when a
small amount of strong acid or base added to it.
the Henderson-Hasselbalch equation, which
describes pH in terms of pKa


[A] is the concentration of the conjugate base
and [HA] is the concentration of the acid.
Determination of pKa values:
pKa values can be obtained from the titrat
ion
The pH at the point of inflection is
the pKa value and this may be read
directly
By definition the pKa value is equal to the
pH at which the acid is half titrated.
compound showed maximum buffering
capacity at its pKa value.

Buffer capacity, , - Quantitative measure of the resistance of a
buffer solution to pH change on addition of hydroxide ions.
Number of moles of acid /base required/ unit change of pH
Buffer value maximum at pKa




where dn - infinitesimal amount of added base and d(p[H+]) is the
resulting infinitesimal change in the logarithm of the hydrogen ion
concentration.
Exp: Prepare 100ml of 0.5M phosphate buffer of pH 6.5 (pKa=7.8)
and determine buffering capacity.
Use the Henderson Hasselbalch equation to find the ratio of
A- to HA.


pH = pKa + log [A-] / [HA]
6.50 = 7.20 + log [A-] / [HA]
-1.3= log [A-] / [HA]
1.3= log [HA] / [A-]
antilog1.3= [HA] / [A-]
19.95 /1 = [HA] / [A-] = KH2PO4 / K2HPO4
KH2PO4 + K2HPO4 = 19.95 + 1 = 21.95
KH2PO4 = 19.95/20.95 X 0.5 = 0.476 M
K2HPO4 = 1/20.95 X 0.5 = 0.0238 M
Moles = wt/MW , wt = moles x MW
wt(KH2PO4)= 136 x 0.476 = 64.736g/l = 6.47g/l
wt (K2HPO4)= 174 x 0.0238 = 4.1412g/l = 0.4141g/l

Titration
Titration curves - by monitoring the pH of
given volume of a sample solution after
successive addition of acid or alkali
Titration curves- Plots of pH vs volume of
titrant added or more correctly against the
number of equivalents added per mole of
the sample
Titration of acid --COOH
At the starting point the acid form
predominates (-COOH).
As strong base is added (e.g. NaOH), the
acid is converted to its conjugate base.
At the mid point of the titration, where pH=pK,
concentrations of the acid = conjugate base
At the end point(equivalence point), the
conjugate base predominates
Total amount of OH added = amount of acid
that was present in the starting point.
Titration
Determination of pKa values
pKa values can be obtained from the
titration data :
1. The pH at the point of inflection is the
pKa value and this may be read directly
2. By definition the pKa value is equal to the
pH at which the acid is half titrated. The
pKa can therefore be obtained from the
knowledge of the end point of the titration.
Titration of glycine
When an amino acid is dissolved in water
it exists predominantly in the isoelectric
form.
Upon titration with acid, it acts as a base,
and upon titration with base, it acts as an
acid( a compound that can act as either an
acid or a base is known as an amphoteric
compound).

+
H
3
N-CH
2
-COO
-
+ HCl
+
H
3
N-CH
2
-COOH + Cl
-
(base) (acid) (1)

+
H
3
N-CH
2
-COO
-
+ NaOH

H
2
N-CH
2
-COO
-
+ Na
+

+H
2
O
(acid) (base) (2)
In this experiment, the amino acid represents
either the A
-
or the HA form in the Henderson-
Hasselbalch equation, depending on the
titration.


Acidbase properties
All of the amino acids have an acidic
group (COOH) and a basic group (NH2)
attached to the carbon.
Two of the amino acids have acidic side
chains: aspartate and glutamate.
Three of the amino acids have basic side
chains: arginine, histidine, and lysine.
All amino acids contain ionizable groups
that act as weak acids or bases, giving off
or taking on protons when the pH is
altered.

These ionizations follow the Henderson-
Hasselbalch equation:
pH=pKa+log [unprotonated form(base)]
[protonated form (acid)]


When the conc of the unprotonated form
equals that of the unprotonated form, the
ratio of their concentrations equals 1, and log
1=0.
Hence, pKa can be defined as the pH at
which the concentrations of the protonated
and unprotonated forms of a particular
ionizable species are equal.
The pKa also equals the pH at which the
ionizable group is at its best buffering
capacity; that is the pH at which the solution
resists changes in pH most effectively.
Consider applying the Henderson-
Hasselbalch equation to the titration of
glycine with acid and base.
Glycine has two ionizable groups: a
corboxyl group and an amino group, with
pKa values of 2.4 and 9.6 respectively.
In water at pH 6, glycine exists as a
dipolar ion, or zwitterion, in which the
carboxyl group is unprotonated(-COO
-
)
and the amino group is protonated to give
the substituted ammonium ion(-NH
3
+
).
Addition of acid to the solution lowers the pH
rapidly at first and then more slowly as the
buffering action of the carboxyl is exerted.
At pH 2.4 the pKa is reached, one-half the
acid has been consumed, and the carboxyl
group is half ionized and is most effective as
a buffer.
Titration of the amino group with base follows
a similar curve into the alkaline region.
The intersection between the titration of the
carboxyl group and the titration of the amino
group describes in this case the point at
which glycine has no net charge, and is
called the isoelectric point (pI).
The isoelectric point (pI)
the isoelectric point, pI, is the pH of an aqueous
solution of an amino acid at which the molecules
have no net charge. In other words, the positively
charged groups are exactly balanced by the
negatively charged groups.
For simple amino acids such as alanine, the pI is
an average of the pK
a
's of the carboxyl (2.34) and
ammonium (9.69) groups. Thus, the pI for alanine
is calculated to be: (2.34 + 9.69)/2 = 6.02.
If additional acidic or basic groups are present as
side-chain functions, the pI is the average of the
pK
a
's of the two most similar acids.
Cont.. (pI)
In the case of aspartic acid, the similar
acids are the alpha-carboxyl function (pK
a

= 2.1) and the side-chain carboxyl function
(pK
a
= 3.9), so pI = (2.1 + 3.9)/2 = 3.0.
For arginine, the similar acids are the
guanidinium species on the side-chain
(pK
a
= 12.5) and the alpha-ammonium
function (pK
a
= 9.0), so the calculated pI =
(12.5 + 9.0)/2 = 10.75.

Most amino acids contain carboxyl and
amino groups having pKa values similar to
those of glycine.
In addition to these groups, many amino
acids contain other ionizable groups,
which introduce other steps or pKa
values into their titration curves.
Titration curves
The pK is the pH at the midpoint of the buffering
region (where the pH changes only slightly upon
addition of either acid or base).
The pK is the pH corresponding to the inflection
point in the titration curve.
The end point of a titration curve represents the
observed end of the titration.
The isoelectric point (isoelectric pH; pI) is the pH
at which the amino acid has a net zero charge. For
a simple diprotic amino acid, the pI falls halfway
between the two pK values. For acidic amino
acids, the pI is given by (pK
1
+ pK
2
) and for
basic amino acids its given by (pK
2
+ pK
3
)


Objective: To determine the pKa and pI value of glycine
amino acid
Take 25 mL of a 0.2 M amino acid solution .
Titrate the amino acid with 1.0 M HCl (titrant)
a. Determine the pH of the amino acid solution before the
addition of titrant.
b. Initially add approximately 0.5 ml of the titrant to the amino
acid at a time.
ml 1.0 M HCl pH
0.0
0.5
In the beginning, the pH will change very dramatically with each
addition of titrant. As you get closer to the pKa of the ionizable
group, the pH will change much more slowly. When this
phenomenon occurs, add 1 ml of titrant at a time.
c. After the addition of each volume of HCl, stir the solution briefly.
d. Turn the stirrer off and measure the pH using the pH meter.
e. Continue with the titration until the pH ~1.5.
3. Repeat steps 1 and 2 above, this time using 1.0 N NaOH as the
titrant for a fresh 25-mL sample of the same amino acid. Record the
data until to pH ~13.
Construct your titration curve plotting pH versus ml of acid and base
added to the amino acid and determine pKa and pI.