Application of Acid base titrations

 Routinely used in industry to analyze products (also helps identify fat and water content
and the presence of vitamins) to be sold. Many manufacturers are under strict standards
of quality control because their products are sold for public consumption.
 It is used for analyzing samples of blood and urine to determine the concentration of
certain chemicals.
 New drugs also owe their existence to the process of titration.
 also applicable in agriculture, soaps, explosives etc

1. Determination of Nitrogen
Kjeldahl Analysis:
 Important method to accurately determine nitrogen in proteins and other nitrogen
containing compounds. (proteins, amines, organic compounds)
 Official worldwide standard for the determination of nitrogen since 1883.
 Also employed in environmental analysis and the agricultural industry for the
determination of nitrates and ammonium.
 Consists of 3 major steps: digestion, distillation and titration.
Steps
i. Digest the material in sulfuric acid convert to NH 4HSO4 decomposes the organic
substance by oxidation to liberate the reduced nitrogen as ammonium sulphate. potassium
sulphate is added to increase the boiling point of the medium. Chemical decomposition of
the sample is complete when the initially very dark-coloured medium has become clear
and colourless.

ii. Cool the solution and distilled with excess base sodium hydroxide, which converts the
ammonium salt to ammonia.Volatile ammonia distilled into known volume of acid (boric
acid, HCl, sulphuric)
iii. Excess acid (boric acid H3BO3) back titrated. The amount of ammonia present, and thus
the amount of nitrogen present in the sample, is determined this way. The ammonia reacts
with the acid and the remainder of the acid is then titrated with a sodium carbonate
solution by way of a methyl orange pH indicator.
Degradation: Sample + H2SO4 → (NH4)2SO4(aq) + CO2(g) + SO2(g) + H2O(g)
Liberation of ammonia: (NH4)2SO4(aq) + 2NaOH → Na2SO4(aq) + 2H2O(l) + 2NH3(g)
Capture of ammonia: B(OH)3 + H2O + NH3 → NH4+ + B(OH)4−
Back-titration: B(OH)3 + H2O + Na2CO3 → NaHCO3(aq) + NaB(OH)4(aq) + CO2(g) +
H2O
The results can be expressed in % N, % NH3 or protein (%N x factor)
In practice, this analysis is largely automated; specific catalysts accelerate the
decomposition
Example

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A protein contains 16.2 wt% nitrogen. A 0.500 mL sample of the protein solution was digested
and the liberated NH3 distilled into 10.00 mL of a 0.02140 M HCl solution. The unreacted HCl
required 3.26 mL of a 0.0198 M NaOH. Calculate the concentration (mg/mL) of protein
in the original sample.

Titration amino acids

Objectives
 To study the titration curves of amino acid.
 To determine the pKa values.
 To determine isoelectric point (pI).
 To determine buffering regions.

Properties of amino acids

 Amino acids are Amphoteric which means it can react as an acid (donate a proton) as
well as a base(accept a proton)
 Amphoteric properties of amino acids are due to the presence of their ionizable α-amino
and α-carboxylic group can act sometimes as acids and sometimes as bases depending on
the pH of their media.
 Each amino acid have a different isoelectric point (pI). pI is the pH value at which the
positive charge equals the negative charge (i.e. the net charge of this molecule equals
zero) (zwitterion)

At low pH, the amino acid is protonated at both the amine and carboxyl functions. At this pH it
carries a net positive charge and can be treated as a diprotic acid, an acid with two pKa's.

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At high pH, both the carboxyl and amine groups are deprotonated. At these pH values, the
amino acid carries a net negative charge, and is dibasic.

At some intermediate pH, the amino acid is a zwitterions, and carries no net charge. This is
called the isoelectric point of the amino acids, and is designated pHI.

By titrating an amino acid and ploting its curve, we can get important information about amino
acid, for example

 pKa and also the pI. Amino acids have more than one pka, because it is polyprotic
(contain more than one ionizable groups).
 Also it provides information about the buffering range of the amino acid that is studied.
 Based on the number of plateaus on a titration curve, one can determine the number of
dissociable protons.

Titration curves are produced by monitoring the pH of given volume of a sample solution
after successive addition of acid or alkali.
The curves are usually plots of pH against the volume of titrant added or more correctly
against the number of equivalents added per mole of the sample
All amino acids contain ionizable groups that act as weak acids or bases, giving off or
taking on protons when the pH is altered.
When an amino acid is dissolved in water it exists predominantly in the isoelectric form.
When this dissolved amino acid is titrated with acid, it acts as a base, and with base it acts
as an acid.
As more of the strong base (titrant) is added to the aqueous solution, more of the weak
acid is converted to its conjugate base. (a buffer forms).
During this process, a buffer system forms and the pH of the system will follow the
Henderson-Hasselbalch relationship.
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, pH=PKa, here, the ionizable group is at its best
buffering capacity; that is the pH at which the solution resists changes in pH most
effectively.
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.

Some amino acids have other ionizable groups in their side chains (R) and these can also
be titrated.

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Let's titrate a solution of the amino acid, Glycine with NaOH

 We begin our titration at a low enough pH (below 2.0) to insure that the amino acid is
fully protonated.

Glycine is a diprotic amino acid which means that it has two dissociable Protons, one on
the α amino group and the other on the carboxyl group.
In Glycine, the R group does not contribute a dissociable Proton.

The dissociation of proton proceeds in a certain order which depends on the acidity of the proton:
the one which is most acidic and having a lower pKa will dissociate first.  So, the H+ on the α-
COOH group (pKa1, =2.34) will dissociate before that on the α-NH3 group (pKa2 =9.69).

 The strong positive charge on the amino group induces a tendency for the carboxylic acid
group to lose a proton, so amino acids are considered to be strong acids.
 These values are typical of pKa's of the carboxyl and amine groups on the α carbon.
 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.
 The titration curve is shown below

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EDTA Titrations
They are Complexation Titrations : based on the reaction of a metal ion with a chemical agent to
form a metal-ligand complex.
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 (chemical
compound in which metallic and nonmetallic, usually organic, atoms are combined.)
Characterized by a ring structure in which a metal ion is attached to two or more nonmetal ions
by covalent bond.
 Metal – Lewis Acid or Electron-pair acceptor
 Ligand – Lewis Base or Electron-pair donor

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Metal ions in solution are always solvated, i.e. a definite number of solvent molecules (usually
2,

4 or 6) are firmly bound to the metal ion.

However, these bound solvent molecules are replaced by other solvent molecules or ions
(ligands) during the formation of a metal complex or metal coordination compound.

 Most chelating agents contain N or O, Elements that contain free electron pairs that
may be donated to a metal.
 Essentially there is formation of coordinate bond…. bond strength between the ligand
and the metal ion is somewhere between that of ionic bonds and covalent bonds
 Metal ions can form complexes with 4, 6, or even 8 ligands at the same time.
 On the other hand the ligand itself may have one place where it can bond to the metal
(Monodentate) or more (Multidentate, form more stable complexes). Because the
entropy of complex formation favours the binding of one large ligand rather than small
ligands.
 EDTA is multidentate chelating agent (ligand).
 EDTA is the most widely used complexing agent for routine analysis of water
hardness and other applications.
 it forms stable complexes with most metal ions – the exceptions being the group
1 cations. Hence most commonly used chelating agent.
 In practice, the use of EDTA as a titrant is well established.
The Chemistry of EDTA
 EDTA= EthyleneDiamineTetraacetic Acid, H4Y shortened further to “Y”.

 It can coordinate to a central metal using up to 6 electron pairs (2 on N and 4 on COO-).

Always forms 1:1 complexes
 The ability of EDTA to potentially donate its six lone pairs of electrons for the formation
of coordinate covalent bonds to metal cations makes EDTA a hexadentate ligand.
 It has four carboxyl groups and two amine groups that can act as electron pair donors, or
Lewis bases.
 H4Y has low solubility in water
 Na2H2Y 2H2O usually used (i.e. two acid groups neutralized) dissociates to H2Y2-to give a
solution of pH 4-5
 In a complexometric titration the analyte and titrant form a complex ion, and the
 Equilibrium constant is called the formation constant, Kf.

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Metal–EDTA Formation Constants
 To illustrate the formation of a metal–EDTA complex, let’s consider the reaction
between Cd2+ and EDTA.

Cd2+(aq)+Y4−(aq) CdY2−(aq)

Kf is quite large, suggesting that the reaction’s equilibrium position lies far to the right.

Hence the reason it is extensively used in standardization of metal cation solutions

EDTA is a Weak Acid: Besides its properties as a ligand, EDTA is also a weak acid. The fully
protonated form of EDTA, H6Y2+, is a hexaprotic weak acid (ionizes stepwise) with successive
pKa values of

pKa1= 0.0 pKa2= 1.5 pKa3= 2.0 pKa4= 2.68 pKa5= 6.11 pKa6= 10.17

The first four values are for the carboxyl protons, and the remaining two values are for the
ammonium protons.

The species Y4– becomes the predominate form of EDTA at pH levels greater than 10.17. It is
only for pH levels greater than 12 that Y4–becomes the only significant form of EDTA.

Fractional composition of EDTA

 We can calculate the fraction of any species in solution using α equation.
 As the pH increases, more EDTA becomes Y4-

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  At any pH a mass balance requires that the total concentration of unbound EDTA equal
the combined concentrations of each of its forms.

T
To save time and prevent errors, the values of α Y
4–
have been determined. There are different
values of αY4– depending on pH.
Why are we worried about the αY4– value?
The complex formation constant between metals and EDTA are usually given in terms of the
complex of the metal and the Y4- form of EDTA.
At any pH lessthat13, less than 100% of the EDTA in solution is in the Y4- form.
So what do we do if w e aren’t at pH=13 ?
We calculate a:
Conditional formation constant

The formation constant for this equation assumes that EDTA is present as Y4–.
Below pH 10.24, the principal species of EDTA is not Y4-, above pH=12 it’s the predominant
species at pH=13 there is exclusively Y4-

 If the pH is controlled by a buffer, then αY4- is a constant that can be combined with Kf

where Kf´ is a conditional formation constant whose value depends on the pH

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 there are different values of the conditional constant depending on pH.
 This allows us to look at EDTA complex formation as if the uncomplexed EDTA were
all in one form.

at lower pH levels, conditional formation constant becomes smaller, and the complex
becomes less stable
Use of the conditional formation constant
Example: Calculate the concentration of free Fe3+ in a solution of 0.10M FeY- at pH 4.00 and pH
1.00.

 EDTA titrations are usually conducted in alkaline conditions.

 For EDTA to react with a metal ion, the hydrogens attached to the
carboxylate groups must be removed.
 In strongly basic solution, these hydrogens are removed by reaction with hydroxide ion.
In more acidic solutions, metal ions must be able to displace the hydrogens if a
complex is to be formed.
 The ability of EDTA to form a complex is dependent on its form. The most desirable
state is the Y4- form.
 Y4- is present at very low concentrations in acidic solution and does not become
the predominant species until the pH exceeds a value of 10.
 Since the actual complexing species is Y4-, complexes will form more efficiently and be
more stable in alkaline solution.
 EDTA is used as primary standard and the certified reference material CaCO3 can be used
to standardize EDTA or to verify the composition of standard EDTA.
Indicators

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Always necessary to use a complexometric indicator to determine when the end point has been
reached.
Because the color of free indicator is pH-dependent, most indicators can be used only in certain
pH ranges, fixed by a buffer.
The most common indicator is the metal ion indicator; most common are organic dyes such as
Fast Sulphon Black and Eriochrome Black T
 To be useful must bind less strongly than EDTA, most common being Eriochrome Black
T.
 EBT binds to metal ions to give a red color. Upon release of the metal to EDTA, it
becomes blue.
 Color change shows that the indicator has been displaced (usually by EDTA) from the
metal cations in solution when the endpoint has been reached.
 Thus, the free indicator (rather than the metal complex) serves as the endpoint indicator.
Can use ion specific electrodes and/or mercury electrodes. Both of these are more expensive
and time consuming.
Sometimes there is not a strong reaction between EBT and the metal.

This can be overcome by a displacement titration. The solution begins with the Mg2+complexed
with EDTA. The analyte is added (assuming higher binding constant and lower concentration)
and the Mg2+is displaced. The Mg 2+ is titrated with EBT.

A second way to overcome titrations with weak end points is to do a back titration. In a back
titration, excess EDTA is added to the sample solution. The excess is then titrated with a
standard Mg or Zn indicator solution.

Titration Selectivity, Masking and Demasking Agents

 EDTA is a widely applicable complexing agent as it will complex with almost any metal.
This can be a problem if selectivity is desired however.
 EDTA is a very unselective reagent because it complexes with numerous doubly, triply
and quadruply charged cations.

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  Several procedures will help to increase the selectivity:
i. Selectivity can be controlled through pH. Controlling the pH is one major
factor that affects complexation.
 The formation of a metal chelate is dependent on the pH of the reaction
medium.
 In weakly acid solution, the chelates of many metals are completely
dissociated such as alkaline earth metals, whereas chelates of Bi, Fe3+ or
Cr are readily formed at this pH.
 Thus, in acidic solution, Bi can be effectively titrated with a chelating
agent in the presence of alkaline earth metals.
 This method is based upon the differences in stability of the chelates
formed between the metal ions and the chelating agent.
 A mixture of bismuth and lead ions can be successfully titrated by first
titrating the bismuth at pH 2 with xylenol orange as indicator, and then
adding hexamine to raise the pH to about 5, and titrating the lead.

ii. Use of masking and demasking agents:

 Masking agents act either by precipitation or by formation of complexes more stable than
the interfering ion-EDTA complex.
 A masking reagent reacts with one of the species and allows titration of the second. This
can be applied to a simple binary mixture or to a more complex mixture. Examples of
Masking reagents:

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 i.
cyanide ion (very poisonous); this forms stable cyanide complexes with the
cations of Cd, Zn, Hg(II), Cu, Co, Ni, Ag, and the platinum metals, but not with
the alkaline earths, manganese, and lead . Hence these can be determined in the
presence of the above mentioned ions.
ii. sulphate for Pb and Ba
iii. oxalate for Ca and Pb
iv. 8-hydroxy quinoline for many heavy metals.
v. Thioglycerol used to mask Cu by precipitation in the assay of lotions containing
Cu and Zn.
 Demasking: the process in which the masked substance regains its ability to enter into a
particular reaction.
 This enables to determine a series of metal ions in one solution containing many cations.
Permits the successive titration of many metals.
 The cyanide complexes of zinc and cadmium may be demasked with formaldehyde-
acetic acid solution

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