Pharmaceutical Analysie

Introduction to
Pharmaceutical Analysls of
the chemical composition
Analytical Chemistry deals with methods for determining all aspects of chemistry viz
in nearly
sample. Analytical Chemistry is found to be useful toolmanufacturing, metallurgical, and
agricultural, clinical, environmental, forensic, identify the substances
to
used
pharmaceutical chemistry. The techniques of this sciencetheareexact amounts of the identified
which may be present in a material and to determine
substance.
Chemical analysis may be classified into four kinds:
amount of each element in a sample with no
(0 Proximate Analysis: It determines the
concern as to the actual compounds present.
of designated constituents in the
(0) Partial Analysis: It deals with the determination
sample. which we are
of partial analysis in
(ii) Trace Constituent Analysis: Aspecialized instance
components present in very minute
Worried about the determination of specified
quantity
component of the sample.
(iv) Complete Analysis: It determines the proportion of each
Chemical analysis are often classified on the basis of sample size:
() MacroAnalysis: The analysis deals with quantities of 0.1 g or more.
() Meso (semimicro) Analysis: This analysis dealing with
quantities ranging from
0.01 to 0.1 g.
g to
(iti) Micro Analysis: This analysis deals with quantities in the range 0.001
0.01 g.
(iv) Submicro Analysis: This analysis deals with samples in the range 0.0001 gto
0.001 g.
(v) Ultramicro Analysis: This analysis deals with quantities below 0.0001 g.
Major Constituent: A major constituent is one accounting for 1-100%.
Minor Constituent: A minor constituent is existing in this range of 0.01-1%.
Trace Constituent: Atrace constituent is presented at absorption of less than 0.01%.
1,2 PHARMACEUTICAL ANALYSIS AND ITS SCOPE
The pharmaceutical analysis is a branch of chemistry, which deals with the process or
sequences of processes to identify and/or quantify a substance or drug, the component of
pharmaceutical solution or mixture or the determination of the structure of chemical
compound used in the formulation of pharmaceutical product.
Some specific uses of analysis are mentioned as follows:
Ö) The examination of raw material, in-process and finished product.
(Gi) Separation, determination and quantitation of impurity.
fii) To check the strength and concentration of the chemical compound
(iv) Used for identification and characterízation of chemical compound
1.2
 Pharmaceutical Analysis Introduction to Pharmaceutical Analysis
(v) To determine the molecular weight of the chemical compound.
(vi) Used for structural elucidation of synthetic compound.
(vii) Quantitative analysis of air, water and soil samples is carried out to determine the
level of pollution.
(vii) In farming, nature of soil and Ilevel of fertilizer application is analyzed.
(ix) In geology,composition of the rock and soil is carried out.
|1.3 DIFFERENT TECHNIQUES OF ANALYSIS
Analysis is broadly divided into two types:
Qualitative Analysis: It gives information about the identity of atomic and molecular
species or the functional groups in samples. It is used only to determine the presence
and absence of the compound.
Quantitative Analysis: It establishes the relative amount of one or more of the
species (analyte) in numerical terms. It measures the concentration or amount of each
substance in a sample.
Analytical Method
Qualitative and/or quantitative analysis are carried out for one or more analytes using a
specified technique/analytical method.
Classification of Analytical Methods
Various methods of analysis can be broadly classified into two categories; Classical
methods and Instrumental methods.
Classical Methods:
These are based on traditional method of analysis and may be divided as:
(a) Qualitative: Use for identification by colour, odours, boiling points, melting point,
limit test etc.
(b) Quantitative: Use for determination of mass or volume (e.q. volumetric and
gravimetric analysis).
" Volumetric Analysis is used to determine the exact volume of the solution of known
concentration reguired to react completely with the solution of the substance to be
analysed.
() Neutralization acid-base titration: It involves neutralization acid base reaction in
presence of water as solvent.
and base in
() Non-aqueous acid-base titration: It involves reaction in between acid
presence of non-aqueous solvent i.e. organic solvents.
() Conmplexometric titration: It is complex formation reaction. In this titration the
reacting substance reacts with standard solution to form a soluble but very slightly
dissociated complex.
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 1.4 METHODS OFEXPRESSING CONCENTRATION
Asolution: It is a homogeneous mixture composed of two or more substances. In such a
mixture, a solute is dissolved in another substance, known as solvent.
Concentrations: It is ameasure how much of given substance there is mixed with
another substance. Most frequenty the concept is limited to homogeneous solutions, where
it refers to the amount of solute in a substance.
Standard solution: It is a chemical term which describes a solution of known
concentration. The concentration of the solution is normaly expressed in units of moles per
Liter (mo/L). Standard solutions are normally used in titrations to determine the
concentration of a substance in solution.
Percent Concentration:
Many a time concentration expressed in terms of percent (parts per hundred), it can be
expressed as,
Mass of solute
1. Percent w/w= Mass of solution X 100

2. Volume of solute
Percent v/v Volume of solution X 100
Mass of solute
3. Percent w/v:=Volume of solutionX 00
Parts Per Million: It is frequently employed to express the concentration of very dilute
solutions and is expressed as 'PPM".
Mass of solute
PPM = x 106
Mass of solution
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 Pharmaceutlcal Analysis
Introduction to Pharmaceutical Analysis
Parts Per Billion: It is also employed to express the
solutions and is expressed as 'PPB'
concentration of extremely dilute

PPB =
Mass of solute
109
Mass of solution x
Molar Solution: It is one that contains one mole of solute
(molecular weight) per litre (L)
of solution. It is indicated by 'M.
Molarity can be calculated as,
The Molecular Weight (MW) The Molecular Weight (MW)
M= or M =
Litre (L) 1000 mL
Examples:
1. AtwO molar solution of aqueous sodium hydroxide (written as "2M
NaOH (aq)")
means there are 2 moles (MW = 40 x 2 = 80 gm)of NaOH per litre of solution.
2. If one litre of asolution contains 98 a of H,SO., this means that the
concentration is
IM.
3. If one litre of a solution contains (98 x 2 = 196 g) of H,SO4, this means that the
concentration is 2M.
Molal Solution: It is one that contains one mole of solute (molecular weight) per
kilogram (kg) of solution. It is represented by 'm'. Molality of solution is independent of
temperature.
Molality can be calculated as,
m=
The Molecular Weight (MW) or m =
The Molecular Weight (MW)
Kilogram (Kg) 1000 gm
It also can be calculated as,

m =
Weight of solute
Weight of solvent xMW of solvent x 100
Example:
Aone molal solution of aqueous sodium hydroxide (written as "1m NaOH (ag)") means
there are 1moles (MW = 40 gm)of NaOH per kilogram of solution.
Normal Solution: It is one that contains one equivalent weights of solute per litre (L) of
solution.
Example:
A two normal solution of aqueous hydrochloric acid (written as "2N HCI (aq)")
means
there are 2 equivalents of HCl per liter of solution.
Normality can be calculated as,
Number of equivalent weights of solute Number eq. Number meq
Normality = Litre of solution L mL
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 PharmaceutlcalAnalysis
Pharmaceutical Analysis Introductlonto

Normality can also be calculated as,

W
N= EW ×V
where, N= Normality
W= Weight of solute
EW =Equivalent weight of solute
V= Volume of solution
Equivalent weight of substance may get changed accordina the reaction carried out in
between the reacting species, normality may get vary. The equivalent weight is generaly
calculated with determination of the species changes in the resulting reaction.
Formal Solution: Some substance (i.e. NaCl or FeCl3) does not exist in molecular form
but remain in ionic form in solid state as well as in solution form. For such substances
formula weight is used for the preparation of solution. It is represented by 'F. On changing
the volume of formal solutions with temperature, formality of the solution may get change.
Formality can be calculated as,
Weight of solute in gm
Volume of the solution in itre XFormula weight
F =
 4.5 PRIMARY AND SECONDARY STANDARDS
1.5.1 Primary Standard
It is not possible all time to prepare volumetric solution of desired theoretical
concentration. Asolution can be prepared at approximate concentration standardized with
the help of primary standard. The concentration (molaríty/normality) obtained after
standardization is used for further quantitative analysis. Volumetric solutions should not
differ by >10% from the prescribed limit.
Primary standard is highly pure stable compound preferably used for the standardization
of less stable secondary standard compound.
Aprimary standard compound should satisfy the following requirement:
1. It must be readily available with less cost.
2. It should be chemically stable.
3. It should be almost 100% pure and in dry form.
4. The total amount of impurities should not exceed 0.01-0.02%.
5. It should be stable at drying temperature and should be dried before weighing.
6. It should not be hygroscopic.
7. Itshould not be oxidized in presence of air and remain stable in air during weighing.
8. It should not be affected by carbon dioxide.
9. It should be readily soluble under the experimental conditions.
10. The reaction with standard solution should be stoichiometric and practically
instantaneous (i.e. must have high rate of forward reaction than backward reaction).
11. The titration error with it should be negligible.
In practice it is very difficult to obtained ideal primary standard and have to compromise
with above requirements.
Examples:
1. Primary standard used in neutralization titration:
(a) For acidic titrant: Anhydrous sodium carbonate.
(b) For basic titrant: Potassium hydrogen phthalate, benzoic acid, oxalic acid.
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 Pharmaceutical Analysis Introductlon to
Pharmaceutical
2. Primary standard used in redox titration:
Analysls
(a) As oxidants: Potassium dichromate, potassium bromated, potassium iodate.
(b) As reluctant: Oxalic acid, sodium oxalate, arsenic trioxide.
3. Primary standard used in complexometric titration:
Pure metallic zincG, pure metallic magnesium, zinc chloride, calcium chloride.
4. Primary standard used in precipitation titration:
Sodium chloride, potassium chloride, potassium bromide.
1.5.2 Secondary Standard
Asecondany standard is less stable compound generally used for quantitative analysis,
and whose concentration can be found by comparison with primary standard. i.e. a titration
with primary standard is carried out and molarity or normality of secondary standard is
determined. The strength of secondary standard must be determined before actual
quantitative analysis of any pharmaceutical compound.
Examples:
1. Secondary standard used in heutralization titration:
(a) Acidic Titrant: Hydrochloric acid, sulphuricacid
(b) Basic Titrant:Sodium hydroxide,
2. Secondary standard used in non-aqueous titration:
(a) Acidic Titrant : Perchloric acid, p-toluenesulfonic acid, 2,4-dinitrobenzenesulfonic
acid.
(b) Basic Titrant : Tetrabutylammonium hydroxide, sodium acetate, potassium
methoxide, sodium aminoethoxide.
3. Secondary standard used in precipitation titration: Silver nitrate, Ammonium
thiocyanate, Potassium thiocyanate, Potassium cyanate, Mercuric nitrate.
4. Secondary standard used in complexometric titration: Ethylenediamine tetra
acetic acid, lead nitrate.
5. Secondary standard used in redox titration: Potassium permanganate, Potassium
dichromate, Iodine solution, Cerric ammonium sulphate.
1.6 PREPARATION AND STANDARDIZATION OF STANDARD
SOLUTIONS
Solutions of accurately known strength are called standard solutions. A standard solution
contains a known weight of reagent in a definite volume of solution. It is neither always
possible nor is it essential, to prepare volumetric solutions of a desired theoretical molarity. A
solution of approximately desired molarity is prepared and standardised by titration against a
solution of a primary standard.
As the strength of a standard solution may change upon standing, the molarity factor
should be re-determined frequently. Volumetric solutions should not differ from the
prescribed strength by > 10% and the molarity should be determined with a precision of
0.2%.
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 Pharmaceutical Analysis
Introductlon to Pharmaceutical Analysis
Stronger or weaker solutions are prepared and standardised using
amounts of the reagent or by making an exact dilution of a stronger solution.proportionate
solutions prepared by dilution should be re-standardised. Volumetric
The water used in preparing volumetric solutions should comply with the
requirements of
the monograph on purified water. When used for the preparation of unstable solutions such
as potassiumpermanganate or sodium thiosulphate, it should be freshly boiled and cooled.
1.6.1 Oxalic Acid (0.1 M)
Molecular formula: H¿C,0, 2H,0 =126
Molecular weight: 90.03
Preparation: Dissolve 6.3 g of oxalic acid in sufficient water to produce 1000 mL
Standardization: Pipette 25 ml of the 0.1M NaOH and transfer it into aclean conical flask,
to it add 2-3 drops of phenolphthalein indicator and titrate with 0.1N oxalic acid solution
until colour changes from purple to colourless.
Reaction:
H,C,O4(a) + 2NaOHa) ’ Na,C,0At) + 2 HzO0
Factor Calculation:
1mole of oxalic acid 2 moles of sodium hydroxide
1000 mL of 1M HCz04 = 80 g of NaOH
1000 mL of 1M H,C,O4 = 40g of NaOH
1mL of 0.1M H,C,0,.2H,0 0.0040 g of NaOH
1.6.2 Sodium Hydroxide (1M)
Molecular formula: NaOH
Molecular weight: 40
Preparation: Dissolve 40g of sodium hydroxide in sufficient carbon dioxide-free water to
produce 1000 mL Prepared solution store in bottles with well-fitted suitable stoppers which
prevent access to atmospheric carbon dioxide.
Standardization: Weigh accurately about 5 gof potassium hydrogen phthalate, previously
powdered and dried at 120°C for 2 hours, and dissolve in 75 ml of carbon dioxide-free water.
Add 0.1 mL of phenolphthalein solution and titrate with the sodium hydroxide solution until
apermanent pink colour is produced. Volumetric solutions of sodium hydroxide must be re
standardise frequently.
Reaction:
COOK cOOK

+ NaOH + H,0
COOH COONa
Sodium
Potassium hydrogen hydroxide Sodium potassium
phthalate phthalate
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 Pharmaceutical Analysis Introduction to Pharmaceutical Analysis

Factor Calculation:
1 mole of NaOH = 1 mole of potassium hydrogen phthalate
1000 mL of 1M NaOH = 204.2 g of CsHsKO4
1 mL of 1 MNaOH 0.2042 g of CGHsKO,
1.6.3 Hydrochloric Acid (1 M)
Molecular formula: HCI
Molecular weight: 36.5
Preparation: Dilute 85 ml of hydrochloric acid with water to produce 1000mL.
Standardization: Weigh accurately about 1.5 g of anhydrous sodium carbonate, previously
heated at about 270°C for 1 hour. Dissolve it in 100 ml of water and add 0.1 ml of methyl
red solution. Add the hydrochloric acid slowly from a burette, with constant stirring, until the
solution becomes faintly pink. Heat the solution to boil, cool and continue the titration. Heat
again to boil and titrate further as necessary until the faint pink colour is no longer affected
by continued boiling.
Reaction:
Na,CO, + 2HCI 2NaCl + H;cO,
H,CO,’ H,0 + CO,1
Factor Calculation:
2 moles of HCI = 1 mole of anhydrous sodium carbonate
2000 ml of 1M HCl = 105.99 g of Na,CO3
1ml of 1M HCI 0.05299 g of Na,CO3
1.6.4 Sulphuric Acid (0.5 M)
Molecular formula: H,SO4
Molecular weight: 98.08
Preparation: Add slowly, with stirring., 27 mL of sulphuric acid to about 1000 ml on
water, allow to cool 25°C.
Standardization: Weigh accurately about 1.5 g of anhydrous sodium carbonate,
previously heated at about 270°C for 1 hour. Dissolve it in 100 ml of water and add 0.1 mL of
methyl red solution. Add the sulphuric acid slowiy from a burette, with constant stirring, until
the solution becomes faintly pink. Heat the solution to boil, cool and continue the titration.
Heat again to boil and titrate further as necessary until the faint pink colour is no longer
affected by continued boiling.
Reaction:
Na,CO, + H;SO4 Na,SO + H,COs
H;CO; H0 + CO;‘
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 Pharmaceutical Analysis Introduction to Pharmaceutical Analysis
Factor Calculation:
1mole of H,SO, l mole of anhydrous sodium carbonate
1000 mL of 1M H,SO4 105.99 g of Na,CO,
1 mL of 0.5 M H,SO, 0.05299 g of Na,CO3
1.6.5 Nitric Acid (1M)
Molecular formula: HNO,
Molecular weight: 63.01
Preparation: Dilute 63 mL of nitric acid with sufficient water to produce 1000 mL.
Standardization: Dissolve 2 a of anhvdrous sodium carbonate in 50 mL of water and
titrate with the nitric acid solution using methyl orange solution as an indicator until the
solution becomes reddish yellow. Boil for 2 minutes,cool and continue the titration until the
reddish yellow colour is restored.
Reactions:
Na,CO3 + 2HNO3 2NaNO3 + HzCO3
HzCO3 H0 + CO,T
Factor Calculation:
2 moles of HNO, 1 mole of anhydrous sodium carbonate
2000 mL of 1M HNO; 105.99 g of Na,CO3
1mL of1 M HNO, = 0.05299 g of Na,CO