CHAPTER 1

REMARKS AND GENERAL DIRECTIONS

I.

Success as an Analyst
a. The student must have the ff:
i. The ability to follow directions under the supervision of a skilled analyst
ii. A fundamental knowledge of the theory as well as a practical ability in the
application of analytical methods
iii. Must know what reactions are taking place during an analysis and be able
to understand and apply the theory upon which the method is dependent
iv. Must acquire skill of technique, patience, neatness, and accuracy
v. Must not only use common sense in the laboratory but think throughout
each step of the procedure
vi. Analysis is carried out not for the sake of analysis but as a means of
controlling the purity and strength of substances used as medicaments

II.

III.

IV.

V.
VI.

Economy of Time
- Best achieved through a preliminary study of the work to be done, followed by a
plan for its prompt execution.
The following ideas or suggestions will save time and increase accuracy
1. Keep the desk clean at all times
2. Clean all apparatus immediately after using it and put it in the desk
3. Label all solutions, filtrates, and precipitates
4. Keep two or more operations going at one time
5. Utilize all time between operations in making calculations and writing up
experiments.
Cleaning Solution
- Prepared by dissolving 200 g of Sodium dichromate (Na2Cr2O7) in 100
mL of water and then adding 1500 mL of sulfuric acid slowly with stirring.
Wash Bottles
- Usually consist of rubber-stoppered, flat-bottomed flasks of 500- to 1000mL capacity fitted with smoothly bent tubes and a jet made flexible by
means of a rubber joint.
Policeman
- A piece of soft rubber tubing fitted to the end of a glass rod.
Reagents
- Used in quantitative analysis and must be pure.
- Conform to the requirements of USP&NF, should be used in all assays
which have for their purpose the establishment of the strength or quality
of official substances

VII.
Purity and Strength Requirements
The purity and strength of chemicals and drugs of the USP and of the NF are usually
expressed in terms of PERCENT (%).

CALCULATIONS OF RESULTS AND ERRORS

ERRORS
-

Refers to the numerical difference between a measured value and the true value.

MAIN TYPES OF ERRORS

1. Indeterminate errors
- Manifest themselves by slight variations in a series of observations made by the
same observer under identical conditions
2. Determinate errors
- They recur in a constant manner in each of a series of determinations
*Arise from causes such as:
a. Personal Errors made by the individual analyst
b. Errors of Method caused by faulty procedure
c. Apparatus Errors due to poor construction or calibration

ACCURACY AND PRECISION

Accuracy is used to denote the agreement of an experimental result or the agreement of the
mean value of a series of experimental result with the true value, and is usually expressed in
terms of error.
i. Absolute Error, the difference between the mean and the true value.
ii. Relative Error, found by dividing the absolute error by the true value.
Precision is a measure of reproducibility of data within a series of results. Results within a
series which agree closely with one another are said to be precise.

AVERAGE DEVIATION is calculated by finding the differences between individual results and
the mean, regardless of sign, adding these differences, and dividing by the number of
determinations.

i.

Relative average deviation is found by dividing the average deviation by the mean.
Expressed in terms of percentage or as parts per thousand.

STANDARD DEVIATION is the preferred measure of precision.

i.

Relative standard deviation, also known as the coefficient of variation.

RANGE is the difference between the largest and smallest results in a series of measurements
SIGNIFICANT FIGURES are defined as all certain digits of a measurement plus one doubtful
digit.

GENERAL OPERATIONS
Crucibles are made of high-grade porcelains. They withstand high temperature and are
suitable for use in the ignition of most drugs and precipitates. Consists of: fused silica,
alundum, nickel, and platinum.

GOOCH FILTRATION CRUCIBLE is designed for the separation of precipitates by suction

filtration.
-Has a perforated bottom upon which is bedded a mat of asbestos, thus making it
possible to collect, wash, dry, and weigh a precipitate in the same crucible.

Evaporation of Liquids, best carried out in porcelain dishes.

-

Evaporations should, with few exceptions, be carried out on a water bath to avoid
danger of loss of material due to spattering or bumping and to avoid
decomposition.

Transfer of Liquids
-

When transferring liquid from one vessel to another, a guide rod should always
be used

Drying and Ignition of Samples and Precipitates

-

Many substances are directed to be dried under specified conditions before

analysis to correct for absorbed moisture.
Precipitates frequently must be dried previous to ignition.

CONSTANT WEIGHT

Dried to constant weight means that two consecutive weighings do not differ by more than 0.5
mg/g of substance taken for the determination, the second weighing following an additional hour
of drying.
-

If the exact weight of an object is desired, the object is heated at the specified
temperature, cooled, and weighed
The object is then reheated, cooled, and again weighed. If the two weights agree
within 0.2 mg, constant weight is said to have been reached.
If the change in wt. is more than 0.2 mg, the process of heating, cooling, and
weighing is repeated until constant weight is obtained.

DESSICATORS
-

Are a special form of glass vessel, rendered airtight by means of ground contact
surfaces, used to maintain a dry atmosphere for objects that might be affected by
moisture or carbon dioxide. Anhydrous Calcium Chloride, a dehydrating agent
charged on the lower compartment of the dessicator.

ANALYTICAL BALANCE
-

the single-pan balance maintains a constant weight on the beam at all times, and
the object is weighed by displacing its equivalent weight from the beam.

CHAPTER 3

Principles of Titrimetric (Volumetric) Analysis

Titrimetric methods
-

Analytical methods in which the volume of a solution of known concentration

consumed during an analysis is taken as a measure of the active constituents in
a sample being analyzed.
a. Analyte or active constituent - chemical substance being analyzed
b. Titrant- solution of known concentration; usually added by mean of buret

Titration- act of adding and measuring the volume of titrant used in the assay
Indicator- chemical which change color at or very near the point in the titration where
equivalent quantities of analyte and tirant have reacted
* Stoichiometric or equivalence point- theoretical point at which equivalent amounts of titrant
and analyte have reacted
* End point- a change which is made apparent by use of indicators
* Titration - near the end point or is very close to the equivalence point
* Results in drug assays are expressed as w/w%, w/v% or v/v%
* Titrimetric analysis- uses equivalents for calculations
1

equivalent of acid neutralizes exactly 1 equiv of a base, and vice versa. 1 equiv of
oxidizing agent reacts with exactly 1 equiv of a reducing agent, and vice versa.

* Equivalent- quantity of a substance that is chemically equivalent to 1.0079 g of Hydrogen ions

* Gram-equivalent weight (GEW) - weight in grams which is chemically equivalent to 1 gram
atom of hydrogen ions (1.0079 g). It is the weight of the substance in grams which contains,
furnishes, reacts with directly or indirectly, or replaces 1 gram atom of hydrogen ions.
* Gram milliequivalent weight (GmEW) - GEW/1000
* Equivalent- number of GEW in the procedure
* Milliequivalent- number of GmEW involved in the procedure
Normality

- Concentration expressed and is defined as the number of equivalents of solute per liter
(equiv/liter) or (meq/liter) of solution

Molarity
- Concentration of a solution in terms of moles per liter

Molality
- Number of solute per 1000g of solvent

Standardization
- Determination of normality and molarity of a solution
a. Primary standard- use of carefully weighed sample of known purity
b. Secondary standard- use of another standard solution with known
concentration
Standard solution- solution of know molarity or normality
Standard acid solutions used in acidimetry and alkalimetry
- HCl used in the titration of compounds that yield a precipitate
- H2SO4 is preferable for hot titrations
Standard alkali solutions - Sodium hydroxide, potassium hydroxide, and barium
hydroxide

Titer
- Weight of a substance which is chemically equivalent to 1 ml of a standard solution

Volumetric apparatus
- made to deliver definite volume of liquid, burets ang pipet; made to contain definite volume
of liquid, volumetric flasks and graduated cylinders
25 degree Celsius- temperature specified in the USF and NF for volumetric measurements
20 degree Celsius- adopted temperature of national bureau of standard for volumetric
apparatus

Milliliter- one thousandth part of a liter; official unit of capacity

Liter- volume occupied by a kilogram of water weighed in a vacuum at 4C
Burets- graduated glass tubes of uniform bore throughout the whole length(mL)

4 types of chemical reactions used in titrimetry

1.
2.
3.
4.

Neutralization- acid reacts with a base (product- salt and water)

Oxidation-reduction
Precipitation
Complexation

4 requirements to be met if the reaction is to be used for titrimetric analysis

1. Reaction must proceed to completion; chemical equilibrium constant must be equal
or greater than to 108
2. Reaction must proceed to stoichiometric manner
3. A suitable end point detecting device must be available
4. Direct titration; must be rapid and must have a sharp end point
Indicators
- complex organic compounds used to determine points in neutralization process, to determine
hydrogen ion concentration, to indicate a desired change in pH has been effected.
3 theories of change in color of indicators
1. Physicochemical theory- increase in pH causes the appearance of new color, and
a decrease in pH causes the disappearance of color or appearance of a different
color
2. Organic theory- change in color to a change in molecular structure
3. Colloidal theory- change in color depends upon the change in size of the colloidal
particle

Table 3.1
Commonly Used pH indicators
Indicator
Malachite green

pH Range
0.0 2.0

Acid
Yellow

Base
Green

Methyl yellow
Bromophenol Blue
Methyl Orange
Bromocresol green
Methyl Red
Bromocresol purple
Bromocresol blue
Phenol Red
Cresol Red
Thymol Blue
Phenolphthalein
Thymolphthalein

2.9 4.0
3.0 4.6
3.2 4.4
4.0 5.4
4.2 6.2
5.2 6.8
6.0 7.6
6.8 8.2
7.2 8.8
8.0 9.2
8.0 10.0
9.3 10. 5

Red
Yellow
Pink
Yellow
Red
Yellow
Yellow
Yellow
Yellow
Yellow
Colorless
Colorless

Yellow
Blue
Yellow
Blue
Yellow
Purple
Blue
Red
Red
Blue
Red
Blue

Preparation and Standardization

Exercise
no.
3.1

3.2
3.3

Title

Standard

Sample

Indicator

End point

Prepare and
Standardize 1N
HCL
1N Sulfuric Acid

Anhydrous
Sodium
Carbonate- PS
Sodium
Hydroxide- SS
Potassium
biphthalate-PS

HCl

Methyl Red

Faint Pink

H2SO4

Phenolphthalei
n
Phenolphthalei
n

Faint Pink

1N Sodium
Hydroxide

NaOH

Permanent Pink

Method of analysis
(Volumetric)
3.1 Direct Alkalimetry
3.2 -Direct Alkalimetry

Analyte

Titrant

Indicator

End point

HCl
H2SO4

1N NaOH
NaOH

Faint Pink
Faint Pink

3.3-Direct Acidimetric

NaOH

0.1N H2SO4

Methyl red
Phenolphthalei
n
Phenolphthalei
n

CHAPTER 4

Permanent Pink

Acidimetric Analysis
Acidimetry
-

direct or residual titrimetric analysis of bases using an accurately measured

volume of acid, is supplied in the analytical control of a considerable number of
official substances, both inorganic and organic.

Direct Titration
-

introduction of a standard acid solution gradually from buret into a solution of the
base being assayed until chemically equivalent amounts of each have reacted as
shown by some change in properties (such as color) of the mixture.

Residual Titration/ Back Titration

- used whenever the end point of a direct deviates appreciably from the steriochiometric
points for some reason.
- Used when a reaction proceeds slowly or when the substance to be assayed does not
give a distinct sharp endpoint with an indicator by direct titration.

Assay of Potassium Sodium Tartrate

-

The NF requires that Potassium Sodium Tartrate contain not less than 99%
and not more than 102% of C4H4KNaO
Methyl red- methylene blue is used as the indicator, since the boiling process
rids the solution of any carbon dioxide which may affect the equivalence point of
the titration.
Methyl red- Methylene blue- Red violet (acid) to dirty blue (transition state) to
green (alkaline)

Assay of Milk of Magnesia

-

USP Requirements: Not less than 7% and Not more than 8.5%

Assay of Drug Sample

Exercise No.

Title

Method of

Chemical

Method of Analysis

Titration
4.1
4.2
4.3
4.4
4.5
4.6
4.7
4.8
4.5
4.6
4.7
4.8

Sodium Bicarbonate
Sodium Hydroxide
Sodium Salicylate
Tablets
Zinc Oxide
Potassium Sodium
Tartrate
Milk of Magnesia
Methenamine
Ammonium Chloride
injection
Potassium Sodium
Tartrate
Milk of Magnesia
Methenamine
Ammonium Chloride
injection

Direct
Direct
Direct

Reaction
Involved
Neutralization
Neutralization
Neutralization

Acidimetric
Acidimetric
Acidimetric

Residual
Residual

Neutralization
Neutralization

Acidimetric
Acidimetric

Residual
Titration
Residual
Residual

Neutralization

Acidimetric

Neutralization
Neutralization

Acidimetric
Acidimetric

Residual

Neutralization

Acidimetric

Residual
Titration
Residual
Residual

Neutralization

Acidimetric

Neutralization
Neutralization

Acidimetric
Acidimetric

Analyte
Sodium Bicarbonate
Sodium Bicarbonate

Titrant
1N Sulfuric Acid
1N Sulfuric Acid

Sodium Salicylate
Zinc Oxide

0.1N HCL
1N Sulfuric Acid

Indicator
Methyl Orange
Methyl Orange or
Phenolphthalein
Bromophenol Blue
Methyl Orange

Endpoint
Faint Pink
Pink
Pale Green Color
Yellow (hot)
Colorless (cool)

1N Sodium Hydroxide
Potassium Sodium
Tartrate

0.5 N Sodium
Hydroxide

Methyl red-methylene
blue

Green

Magnesium Hydroxide

0.5N Sulfuric Acid

1N Sulfuric Acid

Methyl Red

Yellow

Methenamine

1N Sodium Hydroxide
1N Sulfuric Acid

Methyl Red

Ammonium Chloride

0.1 N Sulfuric Acid

Methyl Red

Disappearance of Violet
Color
Red

CHAPTER 5

Alkalimetric Analysis
ALKALIMETRIC ANALYSIS
-

As a general principle, it is recommended that the normality of the

solution obtained by dissolving the acid sample be approximately the
same as that of the titrant.

* Methyl red or phenolphthalein is used as indicators for inorganic acids.

* Phenolphthalein is frequently used as indicator for organic acids; however,
thymol blue, bromothymol blue, and thymolphthalein are also employed.
* Potentiometric methods are also used in determining the endpoint.
* Direct titration requires only one standard solution, the standard base, and
fewer measurements of volume.
* Residual titration methods are used:
-

Whenever direct titrations are not practicable

Applied to official compounds which react too slowly because of poor
solubility; otherwise a heating process is used or precipitation method is used
to convert the substance for reaction with the standard base.

Assay of Drug Sample

Exercise No.

Title

Method of
Titration

5.1
5.2

Hydrochloric Acid
Diluted Phosphoric
Acid
Boric Acid
2-Phenoxyethanol
Tartaric Acid

5.3
5.4
5.5

Analyte
HCl
Diluted Phosphoric Acid
Boric Acid
2-Phenoxyethanol
Tartaric Acid

Method of Analysis

Direct
Blank

Chemical
Reaction
Involved
Neutralization
Neutralization

Direct
Blank
Direct

Neutralization
Neutralization
Neutralization

Alkalimetric
Alkalimetric
Alkalimetric

Titrant
1N Sodium Hydroxide
1 N Sodium Hydroxide
1 N Sodium Hydroxide
1 N Sodium Hydroxide
1 N Sodium Hydroxide

Indicator
Methyl Red
Thymolphthalein
Phenolphthalein
Phenolphthalein
Phenolphthalein

CHAPTER 6

Alkalimetric
Alkalimetric

Endpoint
Pink
Blue
Pink

Nonaqueous Titrimetric Analysis

Kjeldahl method used to determine other organic compounds containing nitrogen

-

Moisture is to be avoided in using nonaqueous procedures, since water, being weakly

basic, would compete with the weak nitrogen base for the HClO4.
Moisture content: Less than 0.05%
Greater care in the control of temperature during standardization and analysis
procedures must be considered.

Neutralization
-

Neutralization is a reaction between hydrogen ions and hydroxide ions in aqueous

solutions of acids and bases
In dioxane (Neutral solvent), or glacial acetic acid, an acidic solvent, perchloric acid
HClO4, is less protophyllic than HCl, and chlorides can be titrated in aqueous solution
with standard HCl.

End Points
-

Indicators used in neutralization reactions in water are in themselves weak acids or

bases which undergo color changes with changes in pH.
Substances of different acid strength require different indicators to detect the end point
of titration
Acidimetry in Nonaqueous Solvents
Weak bases which do not meet this requirement cannot be titrated with accuracy in
aqueous systems because the solvent, water, competes with the basic species in
solution for the proton of the titrant.
To conduct titrations of weak bases, solvent must be changed to eliminate the competing
chemical reaction of h20 for the proton
Titrimetric analysis of a number of weakly basic substances in glacial acetic acid is
possible using a standard solution of perchloric acid as the titrant.

Solvents
-

Solvents used in acidimetric titrimetry are either neutral or acidic in nature

Neutral solvents acetonitrile, alcohols, chloroform, benzene, dioxane, ethyl acetate

(aprotic or amphiprotic in nature)

o Used for solvency action primarily

o Do not enhance dissociation
Acidic solvents formic, glacial acetic, proprionic acids, acetic anhydride, sulfonyl
chloride
o Used for weak bases and their salts

Titrant
-

Perchloric Acid most valuable and widely used titrant in glacial acetic acid
o Strongest of the known common acids
Hydrogen Bromide also used to a limited extent as titrants

Indicators
-

Crystal violet, methylrosaniline chloride, quinaldine red, alpha-naphtholbenzein,

malachite green
o Indicators for titration of weak bases and their salts
Methyl red, methyl orange, thymol blue
o For stronger bases
Crystal violet gives visual end point with less than 0.1 ml of titrant at potentiometric end
point
Alkalimtery in Nonaqueous Solvents
Weakly acidic substances can be titrated in an appropriate nonaqeuous solvent with a
sharp end point
Organic compounds involved: acid halides, anhydrides, acids, amino acids, enols such
as barbiturates and xanthenes, imides, phenols, sulfonamides, and organic salts of
inorganic acids
Boric acid easily titrated using ethylenediamine as solvent

Solvents
-

Factors which can be considered in choosing a basic solvent are:

o Solubility of the substance to be analyzed
o Relative base strength of the solvent
Strong bases ethylenediaminem n-butylamine, morpholine
Weak bases dimethylformamide and pyridine (used for medium-strength acidic
substances
Sulfonamides which contain an n-alkyl substituent possess low acidity; ethylenediamine
would be the solvent of choice for this
N-phenyl or n-pyridyl-substituted sulfonamides; dimethylformamide preferred solvent
Solvent dimethylformamide + thymol blue only sulfathiazole will react with sodium
methoxide

Titrants

Sodium methoxide and lithium methoxide

o Bases used for the titration of acidic compounds
Potassium methoxide stronger titrant; not used because it may produce a gelatinous
reaction product.
Other base titrants: sodium aminomethoxide(strongest base)
o Sodium triphenylmethane: for weakly acidic compounds such as phenols and
pyrroles
Atmospheric moisture and c02 will interfere with metal alcoholate titrations
Blank should not exceed 0.01 ml of titrant per ml of solvent used

Indicators
Potentiometric titration currently the best way of determining the quivalence point of a
given chemical reaction in nonaqueous titrimetry

Azo violet indicator of choice in the titration of acids of weak or intermediate strength in nbutylamine solvent

Thymol blue indicator of choice for the determination of intermediate to strong acids in
dimethylformamide solvent

Ethylenediamine solvent for strong acids

In metal alcoholate, azo violet will change in color before thymol blue

O-nitroaniline for titration of very weak acids

Azo violet/thymol blue endpoint: clear blue color

Other indicators thymolphthalein and p-hydroxyazobenzene

Preparation and Standardization

Exercise No.

Title

Standard
Used

Sample

Indicator

Endpoint

Primary

Potassium
Biphthalate

Crystal Violet

Violet color will change

to emerald green

Primary

Benzoic Acid

Thymol blue

Blue endpoint

0.1N Perchloric
acid
0.1 N Sodium
Methoxide

6.1
6.3

Assay of Drug Sample

Exercis
e No.

Title

Method
of
Titratio
n

6.2

Methacholin
e Chloride

Blank
Titration

Neutralizatio
n

Volumetri
c Analysis

Methacholin
e Chloride

6.4

Phenytoin

Direct
Titration

Neutralizatio
n

Volumetri
c Analysis

Phenytoin

Chemical
Reaction
Involved

Method
of
Analysis

Analyte

Titrant
0.1 N
Perchloric
Acid
0.1 N
Sodium
Methoxid
e

Indicato
r

Endp
t

Crystal
Violet

Azo
Violet

CHAPTER 7

Precipitation and Complexation Methods of Analysis

Volumetric Precipitimetry
Class of reactions is dealt with that require formation of insoluble substances or
precipitates to go to sufficient completion to be quantitative in nature.
Determination of Endpoint:
(a) Cessation of precipitation or appearance of turbidity
(b) Use of internal indicators
(c) Instrumental methods potentiometric, amperometric
Indicators:
(a) Ferric ammonium sulfate, ts thiocyanate reacts with silver or mercuric ions present
to form white precipitate of silver or mercuric thiocyante but as the silver/mercury has
precipitated, the thiocyante ion reacts with ferric ammonium sulfate to form red ferric
thiocyante. RED COLOR MARKS THE ENDPOINT OF REACTION.
(b) Potassium chromate, ts forms red precipitate of silver chromate against the
background of white silver chloride
(c) Adsorption indicators
Dichlorofluorescein (DCF), Eosin Y TS, Tetrabromophenolphthalein ethyl
ester (TEE) prepare them fresh. They are used in the analyses of halides by
direct titration with silver nitrate solution. They are weak organic acids that vary in
strength. Endpoint: color of the silver halide precipitate changes abruptly
because of adsorbed indicator anions. Best seen in a diffuse light condition.

Direct Titration Method

Silver and mercury can be converted into soluble salts estimated by direct titration with
standard ammonium thiocyanate solution using ferric ammonium sulfate as indicator.
Solution must be acidified with HNO3 to prevent hydrolysis that ferric slats undergo in
neutral solution. Chlorides must be absent.

Residual Titration (Volhard) Method

Based on complete precipitation method of insoluble silver salts from nitric acid solution
by addition of excess standard silver nitrate solution to a soluble salt, and determination
of amt of silver nitrate solution in excess by residual titration with standard ammonium
thiocyanate solution using ferric alum as indicator. Produces slightly soluble silver salts:
chlorides, bromide, iodide, cyanide, thiocyanate, sulfide, phosphate, arsentaes,
carbonates

Sodium Tetraphenylboron titration

Na(C6H5)4B, will precipitate organic nitrogen compounds like alkaloids, amines,
quaternanry salts, potassium, ammonium, silver ions
Quaternary compounds will react with bromophenol blue to form blue chloroform soluble
complex, to react with tetraphenylboron to form insoluble compound.
COMPLEXATION METHODS
Quantitative analysis of inorganic pharmaceutical product with metal ions like Al, Bi, Ca,
Mg, Zn
Procedures are time-consuming, since they involve precipitation, filtration, washing,
drying or ignition

Complexation reaction:

Complex when a metal ion combines with a molecule which can donate electrons.
chelate If the combining molecule contains two or more groups that donate electrons.
EDTA (H4Y) is a hexadentate that will react with metal ions to form water-soluble, stable
complex, chelate compounds.
Ligand-attachment to metal ions. EDTA has MW of 372.24 in 1/20 M solution. Or 18.612
g in 1 L of solution.

* Direct (Monovalent), Residual (Polyvalent), Ferric Chloride Titration (based on reaction of

sodium fluoride with ferric chloride to form a reasonably stable complex type of Fef6 3-.
Masking
- Determination of metal in presence of another metal. Capable of entering another
complexation reaction (adjusting pH of titration medium)
Triethanolamine
Thioglycols
Potassium cyanide
Ammonium fluoride
Ascorbic acid, citrates and tartrates
Preparation and Standardization

Exercise
No.
7.1
7.2

Title
0.1 N Silver Nitrate
0.1 N Ammonium
Thiocyanate

Standard
Used
Secondary
Seconday

7.10

0.02 M Sodium
tetraphenylboron

Secondary

7.11

0.05 M Disodium Ethylene

diaminetetraacetate

Primary

Sample
Silver Nitrate
Ammonium
Thiocyanate
Sodium
tetraphenylboro
n
Calcium
carbonate

Indicator

End point

HCl
Ferric
ammonium
Sulfate
Potassium
biphthalate

Precipitate
Red-brown color

Hydroxynaphthol
blue

Blue Color

Precipitate

Assay of Drug Sample

Exercise
No.
7.3
7.4
7.5
7.6
7.7
7.8
7.9
7.12

Title
Phenyl Mercuric Nitrate for Mercury
Content
Sodium Lauryl Sulfate for Sodium
Chloride Content
Iopanoic Acid Tablets
Benzyltrimethyammonium chloride
Sodium Chloride
Iodine Content in Povidone-Iodine
Theophylline
Determination of the Bismuth Content
of Glycobiarsol

Analyte
Phenyl Mercuric Nitrate
Sodium Lauryl Sulfate
Iopanoic Acid
Benzyltrimethylammonium
Chloride
Sodium Chloride
Povidone-Iodine
Theophylline
Glycobiarsol

Method of
Titration
Direct

Chemical Rxn
Involved
Precipitation

Method of Analysis

Direct

Precipitation

Volumetric

Direct
Direct
Residual
Blank
Residual
Residual

Precipitation
Precipitation
Precipitation
Precipitation
Precipitation
Complexation

Volumetric
Volumetric
Volumetric
Volumetric
Volumetric
Volumetric

Titrant
0.1 N Ammonium
thiocyanate
0.1 Silver Nitrate
0.05 N Silver Nitrate
Dichlorofluorescein
TS
Silver Nitrate & 0.1
N Ammonium
Thiocyanate
Silver Nitrate with
0.1 N ammonium
thiocyanate
Silver Nitrate and
Ammonium
Thiocyanate
Disodium EDTA and
0.025 M zinc sulfate

Volumetric

Indicator
Ferric ammonium sulfate

Endpoint
Red Precipitate

Potassium Chromate
Tetrabromophenolphthalein
ethyl ester
0.1 N Silver nitrate

Red Precipitate
Yellow precipitate

Ferric Ammonium Sulfate

Red Precipitate

Ferric ammonium Sulfate

Red Precipitate

Ferric ammonium sulfate

Red Precipitate

Dithizone TS, Alcohol,

Acetic acid-ammonium
acetate buffer

Clear Rose Pink Color

Precipitate

CHAPTER 8

Oxidation Reduction Methods

- Simplest type of oxidation reduction is the direct combination of elements.
Oxidation reduction that take place in the official assay processes are, for the most part,
between electrolytes in aqueous solution. Their quantitative value is based upon the fact that
metals, nonmetals, and their ions, under suitable conditions, can be made to undergo a change
in the quantity of electric charge associated with them and that in the change there exists a
simple relationship between quantity of electricity lost or gained and the weight of the reacting
substances.
Faradays Law a change in charge of one is equivalent to the gain or loss of 96500 C
of electricity for each formula weight of element or group of elements involved.
Reducing agent reactant which loses electrons in an oxidation-reduction reaction
Oxidizing agent reactant containing a constituent atom or atoms which are converted
to a lower state of oxidation
* The equivalent weight of a reducing agent is that weight which loses electrons equivalent to
96500 C
* The equivalent weight of an oxidizing agent is that weight which gains electrons equivalent to
1 faraday

Standard Solutions

Oxidizing agents
Ferric ammonium sulfate
potassium permanganate
potassium dichromate
potassium bromated
potassium iodate
potassium ferricyanide
ceric sulphate
Iodine
bromine

Reducing agents
Ferrous ammonium sulfate
oxalic acid
potassium arsenite
titanium chloride
sodium thiosulfate

* Nitric acid does not undergo a single uniform reaction even though conditions are carefully
controlled

Permanganate Methods
-

Potassium permanganate solution can be standardized easily and retain its

concentrations over long periods of time
Reaction is rapid
Serves as an indicator in titrations where it is used; slight excess of permanganate
imparts distinct pink color

Indirect Titration Methods

-

Employed with those compounds that can be converted through chemical reactions
to an equivalent amount of oxalate, which in turn can then be quantitatively oxidized
by permanganate.

Residual Titration Methods

Two types:
o

Titration in which an excess of standard potassium permanganate solution is

employed to oxidize a substance, and the amount in excess is determined
by reduction with:
Excess standard oxalic acid
Excess ferrous ammonium sulfate and back titration with more
standard potassium permanganate
Titration in which an excess of standard oxalic acid solution is added to the
substance and the excess oxalic acid is titrated with standard potassium
permanganate

* Advantage of ferric alum over oxalic acid is that no heating is required to initiate
reaction of the reducing reagent with the permanganate
* Since permanganate-oxalate reaction is run at elevated temperatures, it is not practical
to titrate permanganate directly with a standard oxalate solution.

Ceric Sulfate Titration Methods

-

Ceric sulfate in diluted Sulfuric Acid is a strong oxidizing agent and considerably
more stable than standard permanganate solutions
Ceric sulfate combine many of the advantages of permanganate and dichromate:
o Solutions are stable even on boiling
o React quantitatively with oxalate or arsenite standard
o Cerous ion is colorless and does not obscure the indicator end point
o No interemediate products are formed in the reduction of ceric cerium
o High concentrations of chloride ion are not oxidized by ceric salts, so that
ferrous iron can be determined in the presence of chlorides
o Ferrous phenanthroline ion is a very satisfactory indicator in titrations with
ceric salts

Orthophenanthroline Test solution

-

Dissolves readily in aqueous solutions of ions known as ferroin which have an

intensely red color
Strong oxidizing agents convert the ferrous to a ferric complex which has a slight
blue color
The color change, red to blue, is reversed by reducing agents
Blue oxidized form is very stable to further change by strong oxidizing agents

Assay of ascorbic acid

-

Dichlorophenol-indophenol: standard oxidizing agent in determination of ascorbic

acid content of ascorbic acid tablets and injection, as well as of the ascorbic acid
content in hexavitamin and decavitamin capsules and tablets
No indicator is necessary

Preparation and Standardization

Exercise
No.

Title

Standard
Used

Sample

Indicator

Endpoint

8.1

Potassium
Permanganate

Primary

*Sodium
Oxalate

*Permanganate
Solution

Pale Pink
Color

8.6

of 0.1 N Oxalic
Acid

Secondary

Oxalic
Acid

*Permanganate
Solution

Pale Pink
Color

8.8

0.1 N Ceric
Sulfate

Arsenic
Trioxide

Orthophenanthro
line Solution

Pale Blue

Assay of Drug Sample

Exercise
No.
8.2

Title

Chemical Rxn
Involved
Redox

Method of Analysis

Hydrogen Peroxide Solution

Method of
Titration
Direct

8.3

Cherry Juice for Malic Acid

Indirect

Redox

Volumetric

8.4

Manganese Dioxide, Precipitated

Indirect

Redox

Volumetric

8.5

Titanium Dioxide

Indirect

Redox
(Reduction)

Volumetric

8.7

Sodium Nitrite

Residual

Redox
(Reduction)

Volumetric

8.9

Ferrous Sulfate Tablets

*Residual

Redox
(OxidationReduction)

Volumetric

Analyte
Hydrogen Peroxide
Cherry juice(for malic acid)
Manganese Dioxide
Titanium Dioxide
Sodium Nitrite

Ferrous sulfate
Hydrogen Peroxide
Cherry juice(for malic acid)

Titrant
0.1 N Potassium
Permanganate
0.1 N Potassium
Permanganate
0.1 N Potassium
Permanganate
0.1 N Potassium
Permanganate;
0.1 N Potassium
Permanganate; 0.1
N Oxalic Acid
0.1 N Ceric Sulfate;
Sulfuric Acid
0.1 N Potassium
Permanganate
0.1 N Potassium
Permanganate

Indicator

Volumetric

Endpoint

---Ferric Ammonium Sulfate

Orthophenanthroline
---

Slight blue color

CHAPTER 9

Oxidation Reduction: Iodimetric and Iodometric

Methods

Ascorbic Acid
- Is a fairly strong reducing agent and is determined by simple titration with iodine
solution, which oxidizes the ascorbic acid to dehydroascorbic acid. Iodine is reduced
to Iodide.
* Organic arsenical can be determined as trivalent or pentavalent arsenic.

LIBERATION OF IODINE FROM POTASSIUM IODIDE AND TITRATION WITH SODIUM

THIOSULFATE
-

Substances that are oxidizing agents which cannot be determined by direct titration
are assayed indirectly. In this method, an equivalent amt of iodine produced by the
oxidation of the iodide ion in an acid medium by the substance is determined by a
standard solution of a reducing agent such as sodium thiosulfate.

OXIDATION-REDUCTION WITH 0.1 N BROMINE

-

Bromine is employed as an oxidizing agent in place of iodine in assay of aniline,

phenol, resorcinol.

Preparation and Standardization

Exercise
. No.
9.1

0.1 N Iodine Solution

Standard
Used
Primary

9.3

0.1 N Sodium Thiosulfate

Primary

9.4

0.1 N Potassium Arsenite

Solution
0.1 N Bromine,
Koppeschaars Solution
0.05 M Potassium Iodate
0.1 M Sodium Nitrite
Solution

Primary

9.11
9.14
9.16

Title

Sample

Indicator

Endpoint

Iodine Solution

Starch TS
Starch TS

Primary

Sodium
Thiosulfate
Potassium
Arsenite Solution
Bromine

Appearance of
Blue Color
Disappearance
of Blue color

Primary
Primary

Potassium Iodate
Sodium Nitrite

Starch TS
Starch TS

Starch TS
Starch TS

Assay of Drug Sample

Exercise
No.
9.2

Title
Antimony Potassium tartrate

Method of
Titration
Direct

Chemical Rxn
Involved
OxidationReduction

Method of
Analysis
Volumetric

9.5

Strong Iodine Solution

Direct

OxidationReduction

Volumetric

9.6

Methionine

Blank

OxidationReduction

Volumetric

9.7

Selenium Sulfide

Blank

OxidationReduction

Volumetric

9.8

Sodium Hypochlorite Solution

Residual

Oxidation-reduction

volumetric

9.2

Antimony Potassium tartrate

Direct

OxidationReduction

Volumetric

9.9
9.10
9.12
9.13
9.15

Cupric sulfate
thyroid
phenol
Ethacrynic acid
Potassium Iodide

blank
blank
blank
blank
Direct

Oxidation-reduction
Oxidation-reduction
Oxidation-reduction
Oxidation-reduction
Oxidation-reduction

volumetric
volumetric
volumetric
volumetric
volumetric

Analyte
Potassium tartrate
Strong Iodine
Methionine
Selenium Sulfide
Sodium hypochlorite
Potassium tartrate
Strong Iodine
Methionine
Cupric sulfate
thyroid
phenol
Ethacrynic acid
Potassium iodide

Iodimetry
Iodometry

Titrant
0.1 N Iodine
0.1 N Potassium
Arsenite
Iodine & 0.1 N
sodium thiosulfate
0.05 N Sodium
thiosulfate
Liberated iodine and
0.1 n sodium
thiosulfate
0.1 N Iodine
0.1 N Potassium
Arsenite
Iodine & 0.1 N
sodium thiosulfate
Liberated iodine
with 0.1 N sodium
thiosulfate
0.1 N sodium
thiosulfate
0.1 N sodium
thiosulfate
0.1 N sodium
thiosulfate
0.05 M Potassium
Iodate

Type of
Titration

Titrant

Direct
Indirect

0.1 N Iodine
0.1 N Na2S2O3

Indicator
Starch TS
Starch TS

Endpoint
Blue color

Starch TS

Disappearance of blue
color

Starch TS
Starch ts
Starch TS
Starch TS

Blue color

Starch TS

Disappearance of blue
color

Starch ts
Starch ts
Starch ts
Starch ts
Starch ts

Primary
Standar
d
As2O3
K2Cr2O7

Indicator

Endpoint

Starch TS
Starch TS

Appearance of Blue Color

Disappearance of Blue
Color

CHAPTER 10

Gravimetric Analysis

The Chemical reactions in gravimetric analysis:

a. Reversible Reactions
- Most of the reactions involved in quantitative analysis are of the reversible type.
Three general conditions tend to prevent reversal and lead to completion of a reaction:
(1) The formation of insoluble gas
(2) The formation of a sparingly soluble gas
(3) The formation of very slightly ionized molecules

b. Law of mass action

- the rate of a reaction is proportional to the product of the molecular concentrations of
the reacting.
*at a definite temperature the equilibrium constant is a fixed value for any given reaction
irrespective of the concentration of the substances present.

c. Solubility product principle

- The product of the concentration of the constituent ions in a saturated solution of a
difficulty soluble salt for any given temperature is practically a constant, each concentration
being raised for a power equal to the relative number of ions supplied by bone molecule of the
salt upon dissociating.

Table 10. 1
Solubility Products of Some Important Salts
Substance

Temperature,
Celsius

Aluminum Hydroxide

25

Barium Carbonate

Barium Sulfate

Calcium Carbonate

Calcium Oxalate

Calcium Sulfate

Lead carbonate

25

25

25

25

25

18

Ions involved

3+
Al + 3OH

2
2+

Ba + CO 3
2
2+

Ba + SO 4
2
2+

+
CO 3
Ca
2
2+

C a + C2 O4
2
2+

+
SO 4
Ca
2
2+

P b + CO 3

Solubility Product

1 x 1033

8.1 x 109

1.1 x 1010

8.7 x 109

2.6 x 109

1.9 x 1014

4.0 x 108

Lead sulfate

18

2+
P b

Magnesium Ammonium
Phosphate

25

2+
M g

2
+ SO 4
+
NH 4 +

1.1 x 1013

2.5 x 1011

3
PO 4

Magnesium Hydroxide

18

1.5 x 105

2+

Mg

+ 2OH
50

Magnesium Oxalate

18

2+

Mg

2
C2 O4

8.8 x 10

18

Mercuric sulfide

Mercurous chloride

Silver bromide

18

2+
2

+
Hg
S

25

2+

+
2Cl
Hg

25

A g + Br

1 x 10

1.5 x 1013

7.7 x 1010

10

Silver chloride

Silver iodide

25

25

A g + Cl
+

A g + l

1.5 x 10

1.5 x 1016

12

Silver thiocyanate

d. common ion effect

25

A g + S CN

1.2 x 10

- the equilibrium constant does not change, no matter what the concentration of the
reacting substances may be. The relative concentration of the reacting substances may change,
but there is no change in the equilibrium constant.
Gravimetric methods:
Gravimetric Analysis, the measurement of the weight of a substance in a sample ot calculation
of the weight of a substance in a sample from the weight of a chemically equivalent amount of
some other substances, can be accomplished in various ways.

2 ways to accomplish gravimetric analysis:

Physical method - is the separation by extraction of a constituent in a natural stated and

weighing of the final product.
Chemical method -is te separation by precipitation or other means of a chemical compound to
the analyst and weighing of the final product

General reaction:
A+B=C+D

Example:
NaCl + AgNO3 = AgCl + NaNO3

A chemically equivalent amount of some product can be obtained from a reactant, i.e., the
component of the sample being assayed, by one of the various methods:
(1) It may be precipitated from solution
(2) It may be a decomposition product resulting from ignition of a compound
(3) It may be deposited on an electrode by electrolysis etc.

Determination of Chlorine in a Soluble Chloride

The silver chloride which precipitates is collected on a filter, washed, dried and weighed. Other
substances that form insoluble silver salts must be absent from the sample.

Determination of Sulfate Ion in a soluble sulfate

The sulfate ion in a soluble sulfate may be determined gravimetrically by precipitation as barium
sulfate, the precipitate being collected, dried, ignited weighed

Determination of the phosphate in sodium phosphate solution

The phosphate is precipitated as magnesium ammonium phosphate, washed, dried, ignited to
the pyrophosphate, and weighed

Precipitation

Precipitant

Product

Determination of Chlorine in
a Soluble Chloride

AgNO3

AgCl

Determination of Sulfate Ion

in a soluble sulfate

BaCl2

BaSO4

Determination of the
phosphate in sodium
phosphate solution

H2S

HgS

Exercise
No.

Title

10.1

Sodium Chloride

10.2

Sodium Lauryl
Sulfate

10.3

Mercaptomerin
Na
Sodium
Phosphate

10.4

Method of
Titration

Chemical
Reaction
Involved

Method of
Analysis

Analyte

Titrant

Gravimetric
Precipitation

Gravimetric

Silver
Nitrate
Barium
Chloride
TS

Gravimetric
Gravimetric

Assay of Drug Sample

CHAPTER 11

Ash and Water Determination

Ash Content
- Residue remains after incineration.
- Inorganic matter added to the crude drug for the purpose of adulteration.
Ash Determination
- A basis for judging the identity and cleanliness of a drug and gives information
relative to its adulteration with inorganic matter.
Total Ash
- The residue remaining after incineration.
- Usually contains carbonates, phosphates,
calcium, magnesium, potassium, sodium,
elements, does not necessarily represent all
since ammonium salts, some alkali iodides
converted to carbonates, etc.
Acid-insoluble Ash

Indicator

sulfates, chlorides, oxides, etc., of

aluminum, iron, and other metallic
the inorganic constituents of the drug,
and nitrates, etoc., are volatilized or

Magnesia
Mixture

Part of the total ash which is insoluble in diluted hydrochloric acid.

*The diluted hydrochloride acid dissolves the calcium carbonate, alkali chlorides, etc., leaving
an acid-insoluble residue that consists almost entirely of silica derived from the soil adhering to
the drug.
Very dull-red heat
Dull-red heat
Bright-red heat
Yellow-red heat
White heat

Temperature
500 to 550C
550 to 700C
800 to 1000C
1000 to 1200C
1200
1600C

Residue on Ignition
- The ash content of chemicals is determined by ignition to dull redness in the same
manner as in the determination of the ash content of the crude drug.
- It is economically impracticable to make ash determinations on large samples of
expensive chemicals.
- Neglible is defined as a quantity not exceeding 500 microgram.

Loss on Ignition
- These technique provides a means of determining the percentage of test material
which is volatilized and driven off under the conditions specified.
- Substances which have a definite limitations on the amounts of volatile matter they
will lose when heated: USP calamine, magnesium sulfate, titanium dioxide, zinc
oxide, NF calcium phosphate, tribasic, kaolin, lime, magnesium hydroxide,
magnesium phosphate.

Water Content
- To ensure uniformity in the official drugs, it is necessary that the USP and the NF
specify certain water-content limits in the drug monographs.
- Water content is generally determined by one of six methods:
Gravimetric, for drugs containing no constituents, other than water, volatile at
105C;
Gravimetric, for drugs containing ether-soluble constituents volatile at 105C;
Azeotropic (Xylene USP; Toluene NF);
Titrimetric (Karl Fischer);
Dew point;
Electrolytic hygrometric.
- The water content is also the moisture content of the drug.

Formulas:

As h Content=

Wt of Residue
x 100
Wt of t h e Sample

Insoluble As h=

Wt of Residue After Treatment of Diluted HCl

x 100
Wt of t h e Sample

Residue on Ignition=

Loss of Ignition=

Sulfated As h=

Wt of Residue After Ignition

x 100
Wt of t h e Sample

Wt of C h emical SubstanceWt of Residue After Ignition

x 100
Wt of C h emical Substance

Wt of Residue after treatment of Conc . H 2 SO 4

x 100
Wt of t h e Sample

Gravimetric Method

Moisture Content=

Wt of Sample Before DryingWt of Sample After Drying

x 100
Wt of the Sample Before Drying

Moisture Content=

Wt of Sample Before DryingWt of Ether Sol . Extract

x 100
Wt of Sample B efore Drying

Azeotropic Method

Moisture Content=

Vol of Water Layer

x 100
Wt of Sample

 Karl Fischer Method

Standard Solution Karl Fischer Solution
Karl Fischer Reagent
o Primary Standard Sodium Tartrate
o Secondary Standard Water-Methanol Solution

Moisture Content=

F=

S ( Volof K . F . Reagent )F(Water Equivalent Factor )

x 100
Wt of the Sample (mg)

Wt of Primary Standard
x 100
Vol of K . F . Reagent

CHAPTER 14

Assay of Volatile Oils

Volatile Oils ethereal oils, essential oils, or essences. They are generally complex products
composed of mixtures of compounds of widely variant characteristics:
The most important chemical components of official volatile oils are:

1. Hydrocarbons occasionally acyclic series, such as heptanes and myrcene but more
often isocyclic series.
Ex: pinene, camphene, limonene, bornylene, fenchene, dipentene, sylvestrene,
and phellandrene.
2. Alcohols present in both the free state and in combination with acids as esters
Ex: linalool, geraniol, citronellol, terpineol, borneol, menthol, and santalol
3. Aldehydes examples are benzaldehyde, cinnamic aldehyde, salicyl aldehye, citral,
and citronellal
4. Ketones examples are camphor, carvone, fenchone, thujone, and menthone
5. Phenols examples are anethol, eugenol, carvacrol, safrol, chavicol, and thymol
6. Acids sometimes present In the free state in small quantities
Ex: acetic, propionic, butyric, valeric, benzoic, cinnamic, and hydrocyanic acids.

More often they occur in combinations with alcohols mentioned under 2 as esters, e.g.,
linalyl acetate, bornyl acetate, and menthyl acetate.

7. Sulfur compounds allyl isothiocyanate (mustard oil)

Preparation and Standardization

Exercise
No.
14.1

Title

Standard Used

Sample

Indicator

Endpoint

Alcoholic
Potassium
Hydroxide

Primary

Lead
acetate

Phenolphthalein
TS

Pale Pink

Assay of Drug Sample

Exercise
No.

Title

Method of
Titration

14.2

Peppermint Oil for Total

Esters

Blank

Chemical
Reaction
Involved

Method of
Analysis

14.3
14.4
14.5
14.6
14.7
14.8

Peppermint Oil for Total

Menthol
Benzaldehyde
Caraway Oil
Clove Oil
Cinnamon
Peppermint Spirit

Analyte
Peppermint Oil
Peppermint Oil
Benzaldehyde

Titrant
0.5 N HCl
0.5 N Sulfuric Acid
1 N NaOH

Blank
Blank

Indicator
Phenolphthalein TS
Phenolphthalein TS
Bromophenol
Phenophthalein TS
KOH TS

Endpoint
Faint Pink
Light-green