QUALITY OF A PRODUCT
• Controlled from START to FINISH
• may deviate from the standard (minimal errors that can
occur) analysis are done to check the quality
• quality analysis is checked
• Quality control is integral in all processes in order to
maintain the quality of a product
PHARMACEUTICAL ANALYSIS
• Provides the criteria that are used to judge the quality of
an analysis
• Control of analytical quality
• Control of errors in analysis
• Validation of analytical procedures
• Cross-checking and monitoring of procedures,
methods and products
VALIDATION USP 1225
• Is defined as the verification, by data and analysis, that
the design objectives of a given facility, system,
apparatus, or procedures are reliably fulfilled in routine
operation.
• A planned series of interactive tests and inspection
➢ designed to describe and reduce uncertainty
in an important process.
• Validated product
➢ appropriate scientific means to be uniform
within a lot, consistent between lots, and
meeting design criteria within defined limits.
PROCESS VALIDATION
• A documented program which provides a high degree of
assurance that a specific process will consistently
produce a product meeting its predetermined
specification and quality attributes.
• Pharmaceutical process validation consists of well
documented, written procedures that define processes
which ensure that a specific pharmaceutical
technology is capable of and is attaining that which is
specified in official or inhouse specifications.
• Prospective or Retrospective
RETROSPECTIVE VALIDATION
• Applicable for a product that has been on the market
➢ adequate data is available for evaluation
• Consists of an evaluation of product characteristics
over time
• Evaluation via control charts
➢ Each attribute could be charted giving a visual
display of the batch history.
➢ allows an evaluation of the consistency of the
process.
➢ helpful in identifying problems and as an aid
in setting practical in-house release limits.
• Useful evaluative procedure - relatively large number
of batches (long period of time)
• Gives detailed information on the product performance.
PROSPECTIVE VALIDATION
• Must ALWAYS be performed for a new product during
initial development and production.
• First three production batches - consistency and
accuracy
• Attributes measured reflect the important or critical
characteristics of the process.
• Requires a knowledge of the process.
• Having identified these features, an experimental
design and sampling plan that captures the relevant
measurements is needed.
• Each type of dosage form or product is different and
may require different considerations.
• Statistical analysis of the data is useful - should be
used to aid in an understanding of the data only.
• Hypothesis testing may not be useful - power
considerations.
• Scientific judgement
VALIDATION OF ANALYTICAL PROCEDURE
• Way of performing the analysis.
• Describe in detail the steps necessary to perform each
analytical test
• Demonstrate that it is suitable for its intended purpose
(ICH Guidelines)
Validation of analytical procedures is directed to the most
common types of analytical procedures:
Identification tests
• ensure identity of an analyte in a sample
• normally achieved via comparison of a property of the
sample
Testing for Impurities
• Quantitative test or a limit test
• Accurately reflect the purity characteristics of the
sample
Assay
• Measure the analyte present in a given sample.
➢ assay represents a quantitative measurement
of the major component(s) in the drug
substance.
Drug Product - similar validation characteristics
VALIDATION CHARACTERISTICS
Specificity
• Ability to assess unequivocally the analyte in the
presence of components which may be expected to be
present.
Identification: Ensure the identity of the analyte
Purity Tests: Accurate statement of the content of impurities of
an analyte
Assay: provide an exact result on the content or potency of an
analyte in the sample
ACCURACY
• Expresses the closeness of agreement between the
value which is accepted either as a conventional true
value or an accepted reference value and the value
found.
➢ trueness
• Assessed using a minimum of 9 determinations over a
minimum of 3 concentration levels covering the
specified range.
• Reported as percent recovery by the assay of known
added amount of analyte in the sample or as the
difference between the mean and the accepted true
value together with the confidence intervals.
PRECISION
• Expresses the closeness of agreement (degree of
scatter) between a series of measurements obtained
from multiple sampling of the same homogeneous
sample under the prescribed conditions.
3 LEVELS OF PRECISION
REPEATABILITY (intra-assay):
• Expresses the precision under the same operating
conditions over a short interval of time.
• Minimum of 9 determinations covering the specified
range for the procedure
• Minimum of 6 determinations at 100% of the test
concentration.
INTERMEDIATE PRECISION
• Expresses within-laboratories variations: different days,
different analysts, different equipment, etc.
 REPRODUCILIBILITY:
• Expresses the precision between laboratories.
• Precision should be investigated using homogeneous,
authentic samples.
• The precision of an analytical procedure is usually
expressed as the variance, standard deviation or
coefficient of variation (relative standard deviation) of a
series of measurements.
• Results should be accurate and precise
• In a normal distribution, 68% of the values should fall
within one standard deviation in either side of the mean
• And 95% should fall within two standard deviations of
the mean
• So, standard deviation should be a small percentage of
the mean.
Detection Limit (LOD)
• Lowest amount of analyte in a sample which can be
detected but not necessarily quantitated as an exact
value
Quantitation Limit (LOQ)
• Lowest amount of analyte in a sample which can be
quantitatively determined with suitable precision and
accuracy.
• Parameter of quantitative assays - low levels of
compounds in sample matrices
• determination of impurities and/or degradation
products.
LINEARITY
• The linearity of an analytical procedure is its ability
(within a given range) to obtain test results which are
directly proportional to the concentration (amount) of
analyte in the sample.
• If there is a linear relationship, test results should be
evaluated by appropriate statistical methods, for
example, by calculation of a regression line by the
method of least squares.
• The correlation coefficient, y-intercept, slope of the
regression line and residual sum of squares should be
submitted.
• A plot of the data should be included. In addition, an
analysis of the deviation of the actual data points from
the regression line may also be helpful for evaluating
linearity.
• For the establishment of linearity, a minimum of 5
concentrations is recommended.
• Other approaches should be justified.
RANGE
• Interval between the upper and lower concentration of
analyte in the sample
ROBUSTNESS
• Measure of its capacity to remain unaffected by small,
but deliberate variations in method parameters and
provides an indication of its reliability during normal
usage.
ASSAY VALIDATION
• Method that provides an estimate of assay accuracy
and precision
To challenge the assay procedure, the following steps are
done:
• Preparation of samples
• Analysis of samples
• Calculation of percent relative deviation
• Calculation of percent relative error
• Disposition
• Documentation
A. PREPARATION OF SAMPLES
Prepare a total of 6 validation samples (or 3 pairs for duplicate
determination) as follows:
1. Prepare two samples to contain the nominal product
formula quantity of the substance to be assayed.
2. Prepare two samples to contain an amount equal to the
nominal product formula minus 1.5 times the
difference between the nominal formula quantity and
the lower assay specification limit.
3. Prepare two samples to contain an amount equal to the
nominal product formula plus 1.5 times the difference
between the nominal formula quantity and the upper
assay specification limit.
B. ANALYSIS
• Two different analysts should carry out a single
determination on each of the three drug levels by the
assay method prescribed for the product.
• The analysts should carry out the assay validation
analysis on different days. Results of the analyses
should be reported as percent of formula quantity.
C. CALCULATION
• The percentage relative deviation (D) is a measure of
the precision of the method.
• In using the formula, ignore the negative sign from
subtraction.
• The percentage relative error (E) is a measure of the
accuracy of the method.
• The lower the values for % Relative Deviation and %
Relative Error, the better the method. All values of %
Relative Deviation (DA, DB, and Dc) and the % Relative
Error (EA, EB, and EC) must be below an acceptable
limit.
• It is difficult to set an acceptable limit for the required
accuracy and precision of a method, since there are
variables that depend on the method being used and
the product being tested.
• There is no substitute for knowledge of analytical
chemistry and good scientific judgment.
• Most modern analytical procedures should have an
accuracy and precision in the range of 1 to 2 percent.
• If a value greater than 2.0 (ignoring the sign) is obtained
for (D) and (E) when the monograph limits are 90% and
110%, a problem with the assay method is indicated.
• A value of 1.0% is used if the monograph limits are 95%
and 105%
 D. DISPOSITION
• Unsatisfactory - subject the procedure to appropriate
RE-VALIDATION
review, designed-study, revision or replacement
• Satisfactory - assay method should be left alone • to ensure continuing validity of the cleaning
procedures.
E. DOCUMENTATION
• Written validation profile Some approaches to validation of cleaning procedures are:

VALIDATION OF MANUFACTURING EQUIPMENTS PRODUCT LINE

Facility or equipment validation
• confirmation that the specified process conditions are
reliably fulfilled in a given apparatus.
The validation program consists of several phases
• installation qualification, operational qualification and
actual validation
INSTALLATION QUALIFICATION
• must cover a number of items such as:
• which is based on the assumption that the active
ingredient is the most deleterious contaminant and that
the mix ratio of active to excipient is indicative of
excipient residue levels.
• Prioritization of product lines chosen for validation is
governed by toxicity of the active ingredient and/or
frequency (volume) of production.
DETERGENT RESIDUE LEVELS
• which are assumed to be independent of the product
and are validated by equipment piece and detergent

POST-VALIDATION
1. Utilities connection to determine if they comply to
regulatory requirements and all applicable corporate • Required whenever there is a change in formulation,
standards, processing conditions, analytical methods, cleaning
2. Equipment features and characteristics: description procedures or materials.
and specifications,
3. Sanitation program COMMON CATEGORIES OF TESTS
4. Maintenance and Drawing
CATEGORY I
OPERATION QUALIFICATION
• Analytical procedures for quantitation of major
• Obtain adequate assurance that the equipment when component of bulk drug substances or active
operated by the approved SOP’s does perform its ingredients in finished pharmaceutical products.
assigned limits.
CATEGORY II
1. Applicable SOP’s • Analytical procedures for determination of impurities in
2. Utilization list bulk drug substances or degradation compounds in
3. Process description finished pharmaceutical products.
4. Key process variables • Quantitative assays and limit tests.
5. Test functions – the items to be tested are indicated.
CATEGORY III
VALIDATION
• Analytical procedures for determination of
• consists of the following steps: performance characteristics

1. State objectives CATEGORY IV

2. Indicate procedure
3. Perform tests • Identification tests
4. Evaluate data and draw conclusions

CLEANING VALIDATION

Contamination - critical factor leading to product failure

• Principal concern is for residual contaminants which

could alter or adversely affect a subsequent product
and cannot be detected by ordinary SOP methods

Cleaning validation program consists of three phrases:

• Pre-validation
• Validation
• Re-validation

• Cleaning SOP’s – ALL EQUIPMENT

• Analytical method selected - sensitive to detect
microgram levels of contaminants.
• TLC, HPLC, UV spectrophotometry, Infrared
spectroscopy

3 Phases of Cleaning validation program:

PRE-VALIDATION

• to evaluate the cleaning, sampling and analytical

testing procedures

VALIDATION

• to establish that the cleaning results are repeatedly

acceptable.
INTRODUCTION TO INSTRUMENTAL METHODS OF ANALYSIS
Classical or Instrumental
CLASSICAL METHODS (WET-CHEMICAL METHODS)
• Measurement depends on the chemical properties of
the sample.
• Reagent - Analyte determined by chemical
stoichiometry.
• Separation of components of interest in a sample by
precipitation, extraction or distillation
• Qualitative analyses of the separated components are
performed based on colors, boiling or melting points,
solubility in different solvents, odors, optical activities
or refractive indices of products formed upon treatment
with reagents.
• Gravimetric or volumetric measurements
INSTRUMENTALMETHODS OF ANALYSIS
• Measurement of physical or chemical properties of the
analyte
• Separation of components of complex mixtures are
performed using chromatography and
electrophoretic techniques.
• Quantitative analyses is based on measurements of
physical properties like conductivity, electrode
potential, light absorption or emission, mass-to-charge
ratio and fluorescence.
INSTRUMENTAL METHODS OF ANALYSIS
• Spectrometric, chromatographic, electrochemical, and
other methods
TYPES OF INSTRUMENTAL METHODS OF ANALYSIS
ANALYTICAL INSTRUMENTS
• Instrument - convert information about the physical or
chemical characteristics of the analyte to information
that can be manipulated and interpreted by man.
• Communication device - system under study and
investigator
• To retrieve the desired information from the analyte, it is
necessary to provide a stimulus, which is usually in the
form of electromagnetic, electrical, mechanical, or
nuclear energy
• The stimulus elicits a response from the system under
study whose nature and magnitude are governed by the
fundamental laws of chemistry and physics.
• The resulting information is contained in the
phenomena that result from the interaction of the
stimulus with the analyte.
SIGNAL GENERATOR
• consists of the chemical system interacting with the
stimulus from the energy source which results in the
production of an analytical signal reflecting the
presence and usually the concentration of the analyte.
TRANSDUCER OR DETECTOR
• transforms the analytical signal produced by the signal
generator into an electrical signal
SIGNAL PROCESSOR
• Modifies and “cleans up” the electrical signal to make it
more convenient to interpret
READ-OUT DEVICE
• converts the electrical signal to a form usable to the
analyst.
CALIBRATION OF INSTRUMENTAL METHODS
CALIBRATION
• determines the relationship between the analytical
response and the analyte concentration.
• determined by the use of chemical standards
TYPES OF CALIBRATION PROCEDURE
• Comparison with standards
• External standard calibration
• Standard addition method
• Internal standard method
A. COMPARISON WITH STANDARDS
DIRECT COMPARISON OR TITRATIONS
• property of the analyte is compared with standards
such that the property being tested matches the
standard.
COLORIMETERS
• the color produced as the result of a chemical reaction
of the analyte was compared with the color produced
by reaction of standards.
TITRATIONS
- among most accurate of all analytical procedures
• Analyte reacts with a standardized reagent (titrant) in a
reaction of known stoichiometry.
• Indicated by the color change of a chemical indicator or
by the change in an instrument response.
• Amount of the standardized reagent needed to achieve
chemical equivalence can then be related to the
amount of analyte present.
B. EXTERNAL STANDARD
• Prepared separately from the sample.
• Used to calibrate instruments and procedures when
there are no interference effects from matrix
components in the analyte solution.
• Ideally – 3 or more solutions, calibration process
• Calibration
➢ Accomplished by obtained the response
signal (absorbance, peak height, peak area) as
a function of known analyte concentration.
• A calibration curve is prepared by plotting the data or by
fitting them to a suitable mathematical equation, such
as the slope-intercept form used in the method of
linear least squares.
 • The response signal is then obtained for the sample
and used to predict the unknown analyte concentration
from the calibration curve or best-fit equation
C. STANDARD-ADDTION METHODS
• Are particularly useful for analyzing complex samples
in which the likelihood of matrix effects is substantial.
• One of the most common form of standard-addition
involves adding one or more increments of a standard
solution to sample aliquots containing identical
volumes
• This process is often called “spiking” the sample.
• Each solution is then diluted to a fixed volume before
measurement.
D. INTERNAL STANDARD
• Is a substance that is added in a constant amount to all
samples, blanks, and calibration standards in an
analysis.
• Calibration involves plotting the ratio of the analyte
signal to the internal standard signal as a function of
the analyte concentration of the standards.
• This ratio for the sample is then used to obtain their
analyte concentration from a calibration curve.
SIGNALS AND NOISES
Analytical measurement - Two components
SIGNAL
• carries information about the analyte that is of interest
to the scientist.
NOISE
• made up of extraneous information that is unwanted
because it degrades the accuracy and precision of an
analysis and also places a lower limit on the amount of
analyte that can be detected.
OUT-OF-CONTROL PROCESS
• A process in which variations among the observed
sampling results cannot be attributed to a constant
system of chance causes.
OUT OF SPECIFICATIONS (OOS RESULT)
• A result that falls outside established acceptance
criteria which have been established in official
compendia and/or by company documentation.
OUT OF TRENT (OOT) RESULT
• A time-dependent result which falls outside a
prediction interval or fails a statistical process control
criterion.
STANDARD
• The metric, specification, gauge, statement, category or
physical product sample against which the outputs of a
process are compared and declared acceptable or
unacceptable.
SPECIFICATION
• A list of tests, references to analytical procedures, and
appropriate acceptance criteria that are numerical
limits, ranges, or other criteria for the test described.
• It establishes the set of criteria to which a material
should conform to be considered acceptable for its
intended use.
SYSTEM SUITABILITY TEST
• Used to verify that the test system will perform in
accordance with the criteria set forth in the procedure.
• The tests are based on the concept that the equipment,
electronics, analytical operations, and samples
analyzed constitute an integral system that can be
evaluated as such.

Chemical analyses are affected by two types of noise:

CHEMICAL NOISE

• Arise from a host of uncontrollable variables that affect

the chemistry of the system being analyzed.
• Examples include undetected variations in temperature
or pressure that affect the position of chemical
equilibria, fluctuations in relative humidity that cause
changes in the moisture content of samples, vibrations
that lead to stratification of powdered solids, changes
in light intensity that affect photosensitive materials,
and laboratory fumes that interact with samples or
reagents.

INSTRUMENTAL NOISE

• Is associated with each component of an instrument,

i.E. The source, input transducer, signal-processing
elements, and the output transducer
• Noise that is observed is usually a complex composite
of several sources of noise that cannot be fully
characterized
• An example is environmental noise which is a
composite of different forms of noise that arise from
surroundings.

The signal-to-noise (S/N) ratio is an equation that indicates the

magnitude of an experimental effect above the effect of
experimental error due to chance fluctuations.

INTERNATIONAL COUNCIL FOR HARMONISATION (ICH)

• An international non-profit organization that aims to

develop guidelines via a process of scientific
consensus with regulatory and industry experts working
together.