PHARMACEUTICAL CHEMISTRY 3

Quality Control I
ANALYTICAL CHEMISTRY
Pharmaceutical Analysis

LUIS P. BAZAN, RPh, MAED, MSPHARMACY, Ph.D.

 Outline of Discussion
I.Introduction to Pharmaceutical Analysis
a. Terminologies
b. QC/QA
c. Pharmacopeia
d. Analytical Methods of Analysis
e. Analytical Method Validation
1. Accuracy
2. Precision
3. Linearity
4. Robustness
5. LOD (limit of detection)
6. LOQ (limit of quantification)
 Outline of Discussion
II. General Methods used in Official Pharmaceutical Analysis

A. Volumetric Analysis
1. Neutralization (aqueous and non-aqueous
2. Precipitation
3. Complexation
4. Redox titration
B. Gravimetric Analysis
 Outline of Discussion
III. Methods used in Analysis of Natural Product

1. Analysis of Crude Drugs

2. Ash /Water content determination
3. Extractive Content
4. Constant, Fats and fixed oils tests
5. Assay of Volatile oils
6. Assay of Alkaloids
QA

GMP

QC
 QA
It is the sum total of the
organized arrangements with
the objective of ensuring that
products will be of the
quality required for their
intended use
 GMP
Is that part of Quality
Assurance aimed at ensuring
that products are consistently
manufactured to a quality
appropriate to their intended
use
 QC
Is that part of GMP concerned
with sampling, specification &
testing, documentation &
release procedures which
ensure that the necessary &
relevant tests are performed
& the product is released for
use only after ascertaining it’s
quality
 QA and QC
• QA is the sum total of • QC is that part of GMP
organized arrangements which is concerned with
made with the object of sampling,
ensuring that product will specifications, testing and
be of the Quality required with in the organization,
by their intended use. documentation,and
release procedures which
ensure that the necessary
and relevant tests are
carried out
 QA and QC
• All those planned or • Operational
systematic actions laboratory techniques
necessary to provide and activities used to
adequate confidence fulfill the requirement
that a product will of Quality
satisfy the
requirements for
quality
 QA and QC

• QA is company based • QC is lab based

Supplementary Training Modules on
Good Manufacturing Practice

Analytical Method Validation

WHO Technical Report Series, No. 937,

2006. Annex 4 Appendix 4
 Validation
Objectives
To discuss various aspects of analytical method
validation including:
• Principles of analytical method validation
• Pharmacopoeia methods
• Non-pharmacopoeia methods
• Approaches to analytical method validation
• Characteristics of analytical procedures
 Validation
Characteristics that should be
considered during validation of
analytical methods include:
– specificity
– linearity
– range
– accuracy
– precision
– detection limit
6.1
– quantitation limit
– robustness
Drug quality as our main business
 The Grim Reality

• Poor quality medicines do not meet

official standards for
STRENGTH
QUALITY

PURITY
PACKAGING
LABELING
 The Grim reality

• Poor quality drugs result in serious health

implications including:
treatment failure
adverse effects
increased morbidity
mortality
development of drug resistance
wasted resources
 Drug quality in the world…
• availability of substandard and counterfeit
drugs in disturbing proportion in many low-
income countries
• lack of reliable drug quality assurance
systems in many developing countries
Qualitative and quantitative
pharmaceutical chemistry
 Analytical chemistry

QUALITATIVE

What is in the sample?

QUANTITATIVE

How much is present in

the sample?
 What it is…
• Application of the procedures of qualitative and
quantitative analytical chemistry
– To the analysis and determination of the purity and
quality of drugs and chemicals used in pharmacy
– To the analysis of the chemical constituents found in
the human body whose altered concentrations during
disease states serve as diagnostic aids
– To the analysis of medicinal agents and their
metabolites found in biological systems
 Therefore, QPC has value in…

Manufacturing Human Physiology

Product Formulation PRODUCT PATIENT Biochemistry

Drug Quality Pharmacokinetics
Drug
Process validation Clinical Pharmacy
Quality

DRUG EFFECT

Pharmacodynamics Pharmacology Toxicology

 What is quality…
• Sum of all factors which contribute directly
or indirectly to the safety, effectiveness,
and reliability of the product
• Ensures that drug products are designed
and produced to meet or exceed customer
requirements for effect and safety
Quality control guarantees within
reasonable limits that a drug
product…
• is free of impurities
• is physically and chemically stable
• contains the amount of active
ingredients as stated in the label
• provides optimal release of active
ingredients when the product is
administered
Quality control for quality drugs

• SHEWHART Cycle  PDCA (Plan-

Do-Check-Act) approach
 FACTORS CONTRIBUTING
TO QUALITY PRODUCTS

Personnel
Validated processes
Procedures

Starting materials Equipment

Packing materials
Premises
Environment
Types of analysis
 Based on sample size

• Ultra micro: < 1 mg

• Micro: 1-10 mg
• Semi micro: 10-100 mg
• Macro: 100 – 1000 mg
Based on extent of determination

Proximate
Total amount of a CLASS or
GROUP of active constituents

Ultimate
Amount of a SINGLE CHEMICAL
SPECIES in a sample
Classification of Analysis
Proximate Ultimate

• Total alkaloidal assay • Atropine

of deadly nightshade determination of
(Atropa belladonna) deadly night shade

• Volatile oil content of • Menthol content of

peppermint peppermint
Based on nature of methods

Tritrimetry
CLASSICAL Gravimetry

Spectrometry
INSTRUMENTAL Polarimetry

Water Content
MISCELLANEOUS Ash Content
Acid Value
General principles
in quality control

STANDARDS and
MONOGRAPHS
 The USP/NF
• A book of public pharmacopeial standards
• Contains standards for medicines, dosage
forms, drug substances, excipients,
medical devices, and dietary supplements
 Parts of a Monograph
Halazone

C7H5Cl2NO4S 270.09
Benzoic acid, 4-[(dichloroamino)sulfonyl]-
P-(Dichlorosulfamoyl)benzoic acid [80-13-7]

>> Halazone contains not less than 91.5 per cent and not more
than 100.5 per cent of C7H5Cl2NO4S, calculated on the dried
basis.
 Parts of a Monograph
Halazone -----------------> OFFICIAL TITLE

EMPIRICAL FORMULA
C7H5Cl2NO4S 270.09 MOLECULAR WEIGHT
Benzoic acid, 4-[(dichloroamino)sulfonyl]-
P-(Dichlorosulfamoyl)benzoic acid [80-13-7]

>> Halazone contains not less than 91.5 per cent and not more
than 100.5 per cent of C7H5Cl2NO4S, calculated on the dried
basis.

CAS Registry Number

OFFICIAL DEFINITION
 Parts of Monograph
• Official title
– generic or non-proprietary name of the drug
• Structural formula
– a formula which shows the arrangement of
the elements making up the compound and
how these elements are bonded to one
another such that stability is attained
 Parts of Monograph
• Empirical formula
– the simplest formula of the compoundgeneric
or non-proprietary name of the drug
• Molecular weight
– Overall weight of the compound obtained by
multiplying the atomic weight of each element
(of the compound) by its subscript and then
getting the sum of all the products
 Parts of Monograph
• Chemical Abstracts Service Number
– Code numbers assigned by CAS to the chemical
substance which provides a unique and universal
identifier for the substance. (The numbers do not
have any inherent meaning.)
• Other chemical names
– Include names based on other systems of
nomenclature or naming like IUPAC name by
International Union of Pure and Applied Chemistry
 Parts of Monograph
• Official definition
– A statement of purity and composition of the
compound or drug which has been
predetermined by the Pharmacopeia to be the
purity at which the drug may generally be
regarded as relatively harmless to living
organisms
 Other Parts of Monograph
• Packaging and storage
• Identification
• Loss on drying
• Chapter Number (where a more detailed
discussion of the test can be found)
• Assay
 General principles
in quality control

ACCURACY and PRECISION

 Accuracy and Precision
Accuracy (Validity)
• the degree to which information matches
true or accepted values
• degree to which the results of a
measurement estimates underlying true
value
• an issue pertaining to the quality of data
and the number of errors contained in a
dataset
 Accuracy and Precision
Precision (Reliability)
• degree to which measurements fluctuates
around a central measurement
• Precise attribute information may specify the
characteristics of features in great detail;
however, that precise data--no matter how
carefully measured--may be inaccurate.
• reproducibility or repeatability
Accuracy and Precision
High precision and low accuracy

High accuracy and low precision

Accuracy and Precision
General principles
in quality control

ERRORS
 Indeterminate errors
• manifest themselves by slight
variations in a series of observations
made by the observer under identical
conditions
• intangible and difficult to detect

e.g. differences in the judgement and

skill of analyst
 Determinate errors
• Recur in a constant manner in each of a
series of determinations
• Possible to partially determine their value
and reduce their effect on the final result
 Determinate errors
Arise from:
• Personal errors made by the individual analyst
(e.g. inability to judge color changes sharply 
habitual reading of endpoints too late)
• Errors of method caused by faulty procedure
(e.g. incorrect sampling, contamination of
precipitates, improper selection of indicators)
 Determinate errors
Arise from:
• Apparatus error due to poor construction
or calibration (e.g. inaccuracy in the
calibration of burets or pipets, inequality in
the length of the arms of the balance,
incorrect weights)
Titrimetric analysis
 Titrimetric Analysis
A titration is a method of analysis
determines the precise endpoint of a
reaction and therefore the precise
quantity of reactant in the titration
flask. A buret is used to deliver the
second reactant to the flask and an
indicator or pH Meter is used to
detect the endpoint of the reaction.
Essentials in titration
Analyte

Titrant the active Indicators

constituent to be
analyzed;
the TITRAND

Volumetric
Chemicals
or standard
capable
solution
of changing
of known
at or near
concentration
the endpoint
 Equivalence Point
• Point when added titrant is chemically
equivalent to the analyte in the sample
• aka STOICHIOMETRIC POINT
• Non-observable to the naked eye
 Endpoint
• Experimental approximate of the
equivalence point
• observable
Titrimetric Analysis

Begin by preparing
the buret. It should
be conditioned and
filled with titrant
solution. The analyst
should check for air
bubbles and leaks,
before proceeding
with the titration.
Titrimetric Analysis

Take an initial volume

reading and record it in
the notebook. Before
beginning a titration, the
analyst should always
calculate the expected
endpoint volume.
Titrimetric Analysis

Prepare the solution to be

analyzed by placing it in a clean
Erlenmeyer flask or beaker. If
your sample is a solid, make
sure it is completely dissolved.
Put a magnetic stirrer in the
flask and add indicator.
Titrimetric Analysis
Use the buret to deliver a
stream of titrant to within a
couple of mL of your
expected endpoint. You will
see the indicator change
color when the titrant hits the
solution in the flask, but the
color change disappears
upon stirring.
Titrimetric Analysis

Approach the endpoint

more slowly and watch
the color of the flask
carefully. Use a wash
bottle to rinse the sides of
the flask and the tip of the
buret, to be sure all titrant
is mixed in the flask.
Titrimetric Analysis

Approaching the endpoint,

one may need to add a
partial drop of titrant. This
can be done with a rapid
spin of a teflon stopcock or
by partially opening the
stopcock and rinsing the
partial drop into the flask
with a wash bottle.
Titrimetric Analysis
Make sure you know what the
endpoint should look like. For
phenolphthalein, the endpoint is
the first permanent pale pink.
The pale pink fades in 10 to 20
minutes.

If you think you might have

reached the endpoint, you can
record the volume reading and
add another partial drop.
Sometimes it is easier to tell
when you have gone past the
endpoint.
Titrimetric Analysis

If the flask looks

like this, you
have gone too
far!
Titrimetric Analysis
When you have reached the
endpoint, read the final
volume in the buret and
record it in your notebook.
Subtract the initial volume to
determine the amount of
titrant delivered. Use this, the
concentration of the titrant,
and the stoichiometry of the
titration reaction to calculate
the number of moles of
reactant in your analyte
solution.
Titrimetric Analysis
Titration with a pH meter
follows the same procedure
as a titration with an indicator,
except that the endpoint is
detected by a rapid change in
pH, rather than the color
change of an indicator.
Arrange the sample, stirrer,
buret, and pH meter electrode
so that you can read the pH
and operate the buret with
ease.
Titrimetric Analysis

To detect the
endpoint
accurately, record
pH vs. volume of
titrant added and
plot the titration
curve as you
titrate.
 Types of titration based on chemical
reaction involved
• Neutralization
– Direct and Residual
– Aqueous and Non-aqueous
– Acidimetry and alkalimetry
 Types of titration based on chemical
reaction involved
• Reduction-Oxidation
–Permanganometry
–Iodimetry
–Iodometry
–Cerimetry
 Types of titration based on chemical
reaction involved
• Reactions involving combination of ions
– Volumetric Precipitation
• Volhard Method
• Mohr Method
• Fajans Method
• Gay Lussac method
• Liebig Method
• Complexation
Gravimetric Analysis
 Gravimetric Analysis
• Method based on measurement of weight
of an analyte or a compound containing
the analyte
 When appropriate to use…
Where weights of reactants and products of
chemical reactions are:
– reproducible
– stable
– reflect the presence of constituents which are
important in the establishment of identity
 Precipitation methods
• Based on isolation of an insoluble
precipitate of known composition
Why precipitation?
• Many metallic elements in their ionic forms
react with negative counter ions to
produce stable precipitates.
 Precipitation methods
• Silver ions form stable and highly insoluble
salts with chloride, bromide and iodide.
 Precipitation methods
• Calcium precipitates quantitatively with
oxalate and can be measured reproducibly
at any of three temperature dependent
plateaus as the oxalate, the carbonate and
the oxide.
 Volatilization methods
• Analyte is volatilized, weighed and the loss
is determined.
 An illustration
• The analyte can be bicarbonate or a
mixture of carbonate and bicarbonate. The
total amount of carbonate in whatever
form is found by placing the analyte in a
solution containing an excess of sulfuric
acid.
 An illustration
• This solution is in a flask connected to incoming
nitrogen gas gently bubbled through the solution
and an exit tube first to a drying agent to absorb
aerosolized water and water vapor and then to a
mixture of NaOH and drying agent to absorb the
CO2 and later subsequently produced by the
absorption by NaOH:
 An Illustration
• The tube containing the NaOH on
asbestos and the CaSO4 to absorb the
final water product is pre- and post-
weighed to given the total amount of
carbonate in the sample. Note that the
nitrogen gas acts only as a carrier and
does not take part in any reaction.
 Approximate temperature
equivalents

Very dull red heat

500 – 550 degrees Celsius

Dull red heat

550 - 700 degrees Celsius
 Approximate temperature
equivalents
Bright red heat
800 - 1000 degrees Celsius

Yellow red heat

1000-1200 degrees Celsius

White heat
1200 - 1600 degrees Celsius
 Terms to define
Ignite to Constant Weight
“Ignition shall be continued at 800 + 25
degrees Celsius unless otherwise
indicated until 2 consecutive
weighings do not differ by more than
0.5 mg/g of the substance taken; the
second weighing following an
additional 15-minute ignition period.”
 Terms to define
Loss on Ignition
• Percentage of test material that is
driven off under the temperature
range of 800 + 25 degrees Celsius
Loss on Drying
• Amount of volatile matter of any kind
that is driven off at 110-120 degrees
Celsius
 Terms to define
Drying to constant weight
“Drying shall be continued at 110-120
degrees Celsius unless otherwise
indicated until 2 consecutive
weighings do not differ by more than
0.5 mg/g of the substance taken; the
second weighing following an
additional hour of drying.”
Special methods of analysis
 Gasometric assay
• Measures the volume of gas liberated or
removed under the conditions described in
the assay
 Tests for volatile oils
• Specific gravity
• Polarization
• Refractive Index
 Specific assays
• Ester content
• Alcohol content
• Aldehyde content
• Ketone content
• Phenol content
 Tests for Fats and Fixed Oils

Acid value
• Number of milligrams of KOH required
to neutralize the free acids in 1 gram of
substance
Ester value
• Number of milligrams of KOH required
to saponify esters in 1 gram of
substance
 Tests for Fats and Fixed Oils

Hydroxyl value
• Number of milligrams of KOH
equivalent to the hydroxyl content of 1
gram of substance
Iodine value
• Number of grams of iodine absorbed
by 100 grams of substance
Types of fixed oils based on Iodine
value

120
Drying Linseed, cod

100-120
Semi-drying Cottonseed, sesame

< 100
Non-drying Olive, almond
 Unsaponifiable mattter
• Refers to the substances that are not
saponifiable by alkali hydroxides but are
soluble in ordinary fat solvents
 Ash content determination
• Total ash
• Acid-insoluble ash
• Water-soluble ash
• Sulfated ash
 Water content determination

METHOD I. Titrimetric Method/ Karl-

Fischer Method
• “1 mL of of the reagent is equivalent to
approximately 5 milligrams of water”
 Contents of KFR

Anhydrous Prevents the

pyridine-sulfur
methanol complex
HYDROIODIC ACID
Iodine React with water
Sulfur dioxide
SULFUR TRIOXIDE

Prevents reverse
Pyridine reaction
 Water content determination

METHOD II. Azeotropic Method/

Distillation Method
METHOD III. Gravimetric Method/
Drying/Thermal Method
• Method of choice for chemical
samples (1-2 grams), vegetable drugs
and biologics
 Alcohol content determination

• METHOD I. Distillation Method

• METHOD II. Gas Liquid Chromatography
Sample problems
 Problem 1
• A 0.30 gram sample of sodium
bicarbonate (MW = 84 grams) requires 4
mL of a 0.975 N sulfuric acid. What is its
purity?
• Answer: 109.2%
 Problem 2
•Calculate the magnesium hydroxide content
of milk of magnesia (MW = 58.32 grams) if a
23.0 gram sample dissolved in 50 mL of a 1.1
N sulfuric acid requires 25 mL of a 1.13 N
sodium hydroxide?

%Mg(OH)2=(50mL x 1.1N) – (25mL – 1.13) x 0.02916 x 100

23 g

•Answer: 3.39%
 Problem 3
• A 0.23 gram sample of sodium chloride
was assayed by Volhard method using 50
mL of a 0.101 N silver nitrate and 12 mL of
0.125 ammonium thiocyanate. Calculate
the % sodium chloride

% NaCl = (50mL x 0.101N ) – (12mL x 0.125N) x 0.05844 x 100

0.23 g
• Answer: 90.2%
 Problem 4
• If a 0.46 gram sample of KI yielded 0.7215
grams of AgI precipitate by gravimetry,
what would be the percent purity of KI
(MW of AgI = 234.7745 grams; MW of KI =
166.0045 grams)?
• Answer: 110.90%
 Problem 5
• If a 2-gram sample of cod liver oil required
4.5 mL of 0.02 N NaOH in the titration of
the free fatty acids, would the oil conform
to the official purity requirement (OR: less
than or equal to 1 mL of 0.1 N NaOH for 2-
gram sample)? What is the acid value of
the oil?
• Answer: Yes; 2.52 mg/g
Thank you very much.
Have a nice day!