pharmacy Under the
PROF. ANUP KUMAR ROY
According to the Food and Drug Administration (FDA),
the goal of validation is to:
“establish documented evidence which provides a high degree
of assurance that a specific process will consistently produce a
product meeting its predetermined specifications and quality
WHO: Defines Validation as an action of providing any
procedure, process, equipment, material, activity or system
actually leads to the expected results.
MASTER PLAN (VMP)
Validation master plan (vmp) – it describes the
basic concept of over all site of validation
The vmp addresses process validation, facility
qualification, Analytical method validation and
is to out line the principles involved in the
qualification and validation of facility , define areas
and systems to be qualified and validated and
provide a programme for achieving and
maintaining a validated status
Validation should thus be considered in the
Totally new process
Process and equipment which have been altered
to suit changing priorities
evidence that a
or system will do
what it purports to
do, based upon a
pre-planned series of
scientific tests as
defined in the
• Is employed when an
existing process can
be shown to be in a
state of control by
applying tests on
samples at strategic
points throughout a
process; and at the
end of the process.
All data is collected
of the process until
evidence that a
process does what it
purports to do,
based on review and
analysis of historical
The manufacturing process, in addition to
the individual equipment, must be validated.
The goal is to create a robust manufacturing
process that consistently produces a drug
product with minimal variation that adheres
to quality criteria of purity, identity, and
Some considerations should be exercised when
selecting the process validation strategy.
Extensive sampling and testing should be
performed on the product at various stages, and
should be documented comprehensively.
Upon completion of the review,
recommendations should be made on the extent
of monitoring and the in-process controls
necessary for routine production.
These should be incorporated into the Batch
manufacturing and packaging record or into
appropriate standard operating procedures
The validation protocol is executed.
The production process is broken down into
individual step and evaluated.
All equipment, production environment and the
analytical testing methods to be used should
have been fully validated.
Master batch documents can be prepared.
Using this defined process a series of batches
should be produced.
In using this approach there is always the risk of
having to modify process parameters or
specifications over a period of time.
Concurrent validation may be the practical
approach under certain circumstances. where
the product is a different strength of a
previously validated product with the same ratio
of active / inactive ingredients
when the number of lots evaluated under the
Retrospective Validation were not sufficient to obtain a
high degree of assurance demonstrating that the process
is fully under control
when the number of batches produced are limited (e.g.
o It is important in these cases however, that the systems
and equipment to be used have been fully validated
previously. A report should be prepared and approved
prior to the sale of each batch
o a final report should be prepared and approved after
the completion of all concurrent batches.
Conducted for a product already being marketed and is based on
extensive data accumulated over several lots and over time.
The source of data for retrospective validation should include batch
documents, process control charts, maintenance log books, process
capability studies, finished product test results, including trend
analyses, and stability results. A minimum of ten consecutive batches
produced be utilized.
Batches manufactured for a defined period (minimum of 10 last
Number of lots released per year
Master manufacturing/packaging documents
Current specifications for active materials/finished products
List of process deviations, corrective actions and changes to
Data for stability testing for several batches
Trend analyses including those for quality related complaints
Any major equipment changes after the initial
validation will result in the need for subsequent
In the end, equipment validation will create
specification ranges and tolerances that will be
applied to the normal operation of equipment.
Following are some of the equipments used in
1. Design qualification
2. Installation qualification
3. Operational qualification
4. Performance qualification
MIXER , OR BLENDER DRYER MILLS
1. CHECK EXTRA PADDLE ,
2. VERIFY PADDLE IS
MOUNTED TO SHAFT
3. VERIFY AUTOMATED
3. CHECK FANS
4. VERIFY INLET
2. VERIFY THE
3. FEEDING AND
STEPS MIXER / BLENDER
1. VERIFY APPROVED PURCHASE ORDER
2. MANUFACTURE AND SUPLLIER
3. PHYSICAL DAMAGE
4. REQUIRED UTILITIES
5. INSTALLATION AS PER INSTRUCTIONS PROVIDED IN THE
6. MAINTAINENCE MANUAL & LIST OF CHANGED PARTS
1. Area decontamination: Area decontamination begins with cleaning.
Different antimicrobial agents are used. It should be noncorrosive,
nontoxic, stable, have good residual action and be inexpensive.different
sanitizing agents used are sodium hypochlorite, isopropyl alcohol,
Betadyne, MIKRO QUAT, alkaline glutaraldehyde.
2. Nonviable particulate monitoring: the quantity and size of the particulate
matter in the environment of the enclosures within the aseptic suite are
normally determined by optical particle counters. Air is drawn into a
chamber of the instrument at measured rate. Individual particles in the air
sample are introduced into the focused beam of light within the chamber.
Any particles present will cause the light to scattered at a particular angle.
This light scattering effect is sensed by photo detector tube. With suitable
amplification and rectification of the resulting signal, the quantity and the
size range of the particles present in the sample are displayed on the
A written report should be available after completion
of the validation. If found acceptable, it should be
approved and authorized (signed and dated).
The report should include at least the following:
Title and objective of study
Reference to protocol
Details of material
Programmes and cycles used
Details of procedures and test methods
Results (compared with acceptance criteria); and
Recommendations on the limit and criteria to be
applied on future basis.
Methods should be validated or
Before their introduction and routine use;
Whenever the conditions change for which
the method has been validated,
e.g., instrument with different
Wherever the method is changed and the
change is outside the original scope of the
Phase 1: Pre-Validation Phase covers all activities relating
to product research and development, formulation, pilot
batch studies, scale-up studies, transfer of technology to
commercial scale batches, establishing stability
conditions, storage and handling of in-process and
finished dosage forms, Equipment Qualification,
Installation Qualification, master production documents,
Operational Qualification, Process Capability.
Phase 2: Process Validation Phase designed to verify that
all established limits of the Critical Process Parameters
Validation Maintenance Phase requiring frequent
review of all process related documents, including
validation audit reports to assure that there have
been no changes, deviations, failures,
modifications to the production process, and that
all SOPs have been followed, including Change
Changes in raw materials.
Changes in the source of active raw material
Changes in packaging material.
Changes in the process.
Changes in the equipment.
Changes in the plant/facility.
Variations revealed by trend analysis.
This process consists of establishment of the performance characteristics and the
limitations of the method.
Method performance parameters are determined using equipment that is:
Method validation is required when:
A new method is been developed
Revision of established method
When established methods are used in different laboratories and different analysts etc.
Comparison of methods
When quality control indicates method changes.
Performance characteristics examined when carrying out method validation are;
Limit of detection (LOD) / Limit of quantification (LOQ)
ACCURACY: Accuracy is an exactness of an analytical
method or closeness of true and observed value.
Determination of accuracy:
The accuracy may be determined by application of
the analytical method to an analyte of known purity
(example:-reference standard) and also by comparing
the results of the method with those obtained using an
alternate procedure that has been already validated.
Accuracy is calculated as the percentage of
recovery by the assay of the known added amount of
the analyte in the sample or the difference between
the mean and accepted true value together with
The ICH guidelines recommended to take minimum
of 3 concentration levels covering the specified range
and 3 replicates of each concentration are analyzed
(totally 3*3=9 determinations).
PRECISION: The precision of the procedure is the
degree of agreement among individual test results
when the method is applied repeatedly to the multiple
samplings of a homogeneous sample.
Determination of Precision: The procedure is applied
repeatedly to separate identical samples drawn from
the homogeneous batch of material and measured by
the scatter of individual results from the mean and
expressed as the standard deviation or as the
coefficient of variation (relative standard deviation).
Precision may be the measure of either the degree
of reproducibility or of repeatability of the analytical
method under normal operating conditions.
According to ICH guidelines repeatability should be
assessed using a minimum of 9 determinations covering
the specified range for the procedure.
Precision should be measured for repeatability
(intra-assay precision), intermediate precision, and
SPECIFICITY: ICH document divides specificity into two categories:
Assay/ impurity testsassay is its ability to measure accurately and
ICH defines specificity of an specifically the analyte of interest in the
presence of other components that might be expected to present in the sample
It is the degree of interference from excipients, impurities or degradation
products ensuring that a peak response is due to a single component only i.e., no
Identification test: is demonstrated by the ability to discriminate between
compounds of closely related structures or by comparison to known reference
materials. Use of positive and negative controls is recommended.
Assay and impurity test: is demonstrated by resolution of the two closest eluting
compounds. If impurities are available it has to be shown that the assay is
unaffected by the presence of spiked material.
If impurities are not available the test results are compared to a second well-
Determination of Specificity: When chromatographic procedures are used
representative chromatograms should be presented to demonstrate the degree of
Samples generated by stress testing of the drug substances using acid and base
hydrolysis, temperature, photolysis and mass spectrometry may be useful to show
that the chromatographic peak is not attributable to more than one component
SELECTIVITY: it is a procedure to detect
qualitatively the analyte in the presence of
components that may be expected to be present in
the sample matrix or the ability of a separative
method to resolve different compounds. It is the
measure of the relative location of two peaks.
Determination of Selectivity: Is determined by
comparing the test results obtained on the
analyte with and without the addition of the
potentially interfering material. When such
components are either unidentified or unavailable
a measure of selectivity can be obtained by
determining the recovery of a standard addition of
pure analyte to a material containing a constant
level of the other components.
SENSITIVITY: Sensitivity is the capacity of the test
procedure to record small variation in concentration.
It is the slope of the calibration curve.
LIMIT OF DETECTION: The Limit of detection is the
lowest concentration of the analyte in a sample that
can be detected but not necessarily determined in a
quantitative fashion using a specific method under the
required experimental conditions. Such a limit is
expressed in terms of a concentration of analyte
(example:-mcg/lit ) in the sample.
Measurement: is based on
Signal to noise ratio.
Visual evaluation (relevant chromatogram acceptable)
The standard deviation of the response and the slope.
‘S’ - is the slope of the calibration curve for analyte (
in case of spectrophotometric method).
(In case of chromatography, „S‟ is measured from peak
to peak variation in the baseline signal).
LIMIT OF QUANTIFICATION: The LOQ is the lowest
concentration of analyte in a sample that may be
measured with an acceptable level of accuracy and
precision under the stated operational conditions of the
method. LOQ can vary with the type of method
employed and the nature of the sample.
Measurement: For instrumental and non instrumental
methods the quantitation limit is generally determined
by the analysis of the samples with known
concentration of the analyte
and by establishing the minimum level at which the
analyte can be determined with acceptable accuracy
In case of instrumental methods that exhibit back
ground noise the ICH document describes to compare
measured signals from samples with known
concentration of analyte with those of blank samples.
A typically acceptable signal –to – noise ratio is 10: 1.
LINEARITY AND RANGE:
Linearity is the ability of the method to elicit test results that are
directly proportional to the analyte concentration within a given
Range of an analytical method is the interval between the upper
and lower levels of analyte.
Measurement: - A range of standards should be prepared
containing at least 5 different concentrations of analyte which are
approximately evenly spaced and span 50-150% of the label claim.
At least 6 replicates per concentration to be studied. Plot a graph
of concentration (on X- axis) Vs mean response (on Y- axis).
Calculate the regression equation, Y-intercept and correlation
Plot another graph of Concentration (on X-axis) Vs response ratio
(replicate response divided by concentration, on Y- axis).
The range of the method is validated by verifying that the
analytical method provides acceptable precision, accuracy and
linearity when applied to samples containing analyte at the
extremes of the range as well as within the range.
RUGGEDNESS: Ruggedness is the degree of
reproducibility of test results obtained by the analysis
of the same samples under a variety of test conditions
such s different laboratories , analysts, instruments,
reagent lots, elapsed assay times, temperature , days
It can be expressed as lack of influence of the
operational and environmental variables on the test
results of the analytical method
Determination: By analysis of aliquots from
homogeneous lots in different laboratories by different
analysts using different operational and environmental
conditions that may differ but are still within the
ROBUSTNESS: It is the measure of capacity of an assay
to remain unaffected by small but deliberate variations
in method parameters and provide an indication of its
reliability in normal usuage.degradation and variations
in chromatography columns, mobile phases and
inadequate method development are common causes of
lack of robustness.
Determination of robustness:
Method characteristics are assessed when one
or more operating parameter is varied by
following certain designs.
Full factorial design: consider a six parameter are to
be investigated at two different levels would require
64 (2)6 experiments, which is tedious and not
feasible. So this method is prohibited.
Matrix approach: (Plackett-Burman approach): It is a
saturated fractional factorial design. This reduces
the number of experiments required to study the
effect of the various parameters.( Example; consider
7 parameter 2 level design would require only
8experiments including one dummy)
Re-validation provides the evidence that changes in
a process and the process environment that are
introduced do not adversely affect process
characteristics and product quality.
Documentation requirements will be the same as for
the initial validation of the process. Periodic review
and trend analysis should be carried out at
Validation is used for getting proper
Used for maintaining equipment.
Further convinience and easy to maintain
production process and recheking the result.