Definition

Validation is establishing documented evidence

which provides a high degree oI assurance that a
speciIic process (such as the manuIacture oI
pharmaceutical dosage Iorms) will consistently
produce a product meeting its predetermined
speciIications and quality characteristics.
55ication
The application oI validation will result in
Iewer product recalls and troubleshooting
assignments in manuIacturing operations and
more technically and economically sound
products and their manuIacturing processes.
enefits
A more questioning approach to equipment
and process control and a greater
understanding oI how the process work.
Ability to high light the areas oI protocol
weakness that may be corrected.
Providing the Ioundation Ior eIIective
monitoring and precise in-process control.
Encouraging communication and exchange oI
ideas between diIIerent disciplines.
-ective
To have uniformity and re5roduci-iity of the
5roduct and high quaity.
Vaidation is an eement of the system of quaity
assurance which guarantees for given
5harmaceutica 5roduct.
The attainment of quaity as s5ecified during
routine 5roduction, 5ackaging and contro.
Vaidation 5rocess 5roduce quaity 5roducts with
highest 5ossi-e confidence.
.-
The foowing order of im5ortance or
5riority with res5ect to vaidation is
suggested:
A. SteriIe Products and Their Processes
. Large-voume 5arenteras (LVPs)
2. Sma-voume 5arenteras (SVPs)
3. 5hthamics, other sterie 5roducts, and
medica devices.
. NonsteriIe Products and Their Processes
. ow-dose/high-potency tablets and
capsules/transdermal delivery systems (TDDs)
2. Drugs with stability problems
3. Other tablets and capsules
4. Oral liquids, topicals, and diagnostic aids
Master Plan or Outline of a Process Validation Program
----------------------------------------------------------------------------------------------------------------------------------
Obiective Proving or demonstrating that the process works
Type oI validation Prospective, concurrent, retrospective, revalidation
Type oI process Chemical, pharmaceutical, automation, cleaning
DeIinition oI process Flow diagram, equipment/components, in-process,
Iinished product
DeIinition oI process output Potency, yield, physical parameters
DeIinition oI test methods Method, instrumentation, calibration, traceability,
precision, accuracy
Analysis oI process Critical modules and variables deIined by process
capability design and testing program
Control limits oI critical variables DeIined by process capability design and testing program
Preparation oI validation protocol Facilities, equipment, process, number oI validation trials,
sampling Irequency, size, type, tests to perIorm, methods
used, criteria Ior success
Organizing Ior validation Responsibility and authority
Planning validation trials Timetable and PERT charting, material availability, and
disposal
Validation trials Supervision, administration, documentation
Validation Iinding Data summary, analysis, and conclusions
Final report and recommendations Process validated, Iurther trials, more process design, and
testing.
ethods of Vaidation
. Pros5ective (Pre market) Vaidation
2. Retros5ective Vaidation
3. Re Vaidation
Prospective (Pre market) Validation
It is an experimental plan called the validation protocol is
executed (Iollowing completion oI the qualiIication trials)
beIore the process is put into commercial use.
Most validation eIIorts require some degree oI prospective
experimentation to generate validation support data.
This particular type oI process validation is normally carried
out in connection with the introduction oI new drug products
and their manuIacturing processes.
%0 1ormaliz0d proc088 validation program 8ould n0v0r b0
und0rtak0n unl088 and until t0 1ollowing op0ration8 and
proc0dur08 av0 b00n compl0t0d 8ati81actorilv
. The faciities and equi5ment in which the 5rocess vaidation is to -e
conducted meet CGP requirements (com5etion of installation
qualification)
2. The o5erators and su5ervising 5ersonne who wi -e "running the
vaidation -atch(es) have an understanding of the 5rocess and its
requirements
3. The design, seection, and o5timization of the formua have -een
com5eted
4. The quaification trias using (size) 5iot-a-oratory -atches have
-een com5eted, in which the critica 5rocessing ste5s and 5rocess
varia-es have -een identified, and the 5rovisiona o5erationa
contro imits for each critica test 5arameter have -een 5rovided
5. Detaied technica information on the 5roduct and the manufacturing
5rocess have -een 5rovided, incuding documented evidence of
5roduct sta-iity
6. Finay, at east one quaification tria of a 5iot-5roduction ( size)
-atch has -een made and shows, u5on scae-u5, that there were no
significant deviations from the e5ected 5erformance of the 5rocess.
The o-ective of 5ros5ective vaidation is
to 5rove or demonstrate that the 5rocess
wi work in accordance with a vaidation
master 5an or 5rotoco 5re5ared for 5iot-
5roduct (size) trias.
2. #etrospective VaIidation
Where 5ros5ective vaidation is not ustified for
economic consideration and resources
imitations the retros5ective vaidation is usefu
. it is a55ied to esta-ished 5rocess where data is
avaia-e for statistica and where there has
-een no significant changes in 5rocess, raw
materias or anaytica methods.
2. esta-ishing documented evidence through
review / anaysis of historica manufacturing and
5roduct testing data to verify that s5ecific
5rocess can consistenty 5roduce meetings its
5redetermined s5ecifications and attri-utes.
. Gather the numerical data Irom the completed batch record and include
assay values, end-product test results, and in-process data.
2. Organize these data in a chronological sequence according to batch
manuIacturing data, using a spreadsheet Iormat.
3. Include data Irom at least the last 2030 manuIactured batches Ior analysis.
II the number oI batches is less than 20, then include all manuIactured
batches and commit to obtain the required number Ior analysis.
4. Trim the data by eliminating test results Irom noncritical processing steps
and delete all gratuitous numerical inIormation.
5. Subiect the resultant data to statistical analysis and evaluation.
6. Draw conclusions as to the state oI control oI the manuIacturing process
based on the analysis oI retrospective validation data.
7. Issue a report oI your Iindings (documented evidence).
. oncurrent VaIidation
n-5rocess monitoring of critica 5rocessing ste5s
and end-5roduct testing of current 5roduction can
5rovide documented evidence to show that the
manufacturing 5rocess is in a state of contro.
Such vaidation documentation can -e 5rovided
from the test 5arameter and data sources discosed
in the section on retros5ective vaidation.
Test 5arameter Data source
------------------------------------------------------------------------------
verage unit 5otency
Content uniformity End-5roduct testing
Dissoution time
Weight variation
Powder--end uniformity
oisture content
Partice or granue size distri-ution
Weight variation
Ta-et hardness n-5rocess testing
5 vaue
Coor or carity
Viscosity or density
#evaIidation
Conditions requiring revaidation study and documentation
are isted as foows:
. Change in a critical com5onent (usuay refers to raw
materias)
2. Change or re5acement in a critical 5iece of moduar
(ca5ita) equi5ment
3. Change in a faciity and/or 5ant (usuay ocation or site)
4. Significant (usuay order of magnitude) increase or
decrease in -atch size
5. Sequentia -atches that fai to meet 5roduct and 5rocess
s5ecifications
Ty5es of vaidation
. Ceaning Vaidation
2. Equi5ment Vaidation
3. Personne Vaidation
4. Process Vaidation
5. naytica ethod Vaidation
6. Vendor/su55ier Vaidation
7. Raw materia Vaidation
naytica ethods of Vaidation
ethod vaidation is the 5rocess to confirm
that the anaytica 5rocedure em5oyed for
a s5ecific test is suita-e for its intended
use.
ethods need to -e vaidated or revaidated as
foows:
. efore their introduction into routine use
2. Whenever the conditions change for which the
method has -een vaidated (e.g., instrument with
different characteristics)
3. Whenever the method is changed, and the
change is outside the origina sco5e of the method
4. When quaity contro indicates an esta-ished
method is changing with time
5. n order to demonstrate the equivaence -etween
two methods (e.g., a new method and a standard)
Steps in Method Validation
. Develop a validation protocol or operating procedure Ior the validation.
2. DeIine the application, purpose, and scope oI the method.
3. DeIine the perIormance parameters and acceptance criteria.
4. DeIine validation experiments.
5. VeriIy relevant peIormance characteristics oI equipment.
6. QualiIy materials (e.g., standards and reagents).
7. PerIorm prevalidation experiments.
8. Adiust method parameters or/and acceptance criteria iI necessary.
9. PerIorm Iull internal (and external) validation experiments.
0. Develop SOPs Ior executing the method in the routine.
. DeIine criteria Ior revalidation.
2. DeIine type and Irequency oI system suitability tests and/or analytical quality
control (AQC) checks Ior the routine.
3. Document validation experiments and results in the validation report.
A validation report should be prepared that includes
Description oI the method.
Obiective and scope oI the method (applicability, type).
Summary oI methodology, including sampling
procedures.
Type oI compounds and matrix.
Pro5osed Sequence of Vaidation E5eriments, Eam5e igh
Performance Liquid Chromatogra5hy
-----------------------------------------------------------------------------
Validation parameters Measurement methods
-------------------------------------------------------------------------------
. SpeciIicity with SuIIicient separation oI all
standards compounds (resolution Iactor
~2.5)
2. inearity Iniect Iive standards containing the Iull
working concentrations. Iniect each
standard three times. Average the peak
area. Plot the averaged peak area vs.
concentration. Calculate the linear
regression.
-----------------------------------------------------------------------------
Validation parameters Measurement methods
-------------------------------------------------------------------------------
3. Precision oI the amounts Iniect a standard at three
diIIerent concentrations
Iive times. Calculate
relative standard
deviation oI peak areas.
4. Accuracy
Spike a blank sample with the analyte at three diIIerent
concentrations. Calculate the deviation oI the results
obtained with the method to be validated with the true
value.
5. Intermediate precision
Iniect three
standards at diIIerent concentrations over 5
working days. The analysis should be conducted
by three diIIerent operators using columns Irom
three diIIerent batches. Measure the precision oI
amounts.
6. Limit of detection (LD)
nect a standard with a concentration cose to
the detection imit three times. verage signa height and -aseine
noise. LD = 3 signa height standard amount/-aseine noise
7. Limit of quantitation (L")
S5ecify a 5recision imit for the amount
at the imit of quantitation. Pre5are si standard soutions with the
amounts in the range from the e5ected imit of quantitation to 2
times this amount. nect a sam5es si times and cacuate the
standard deviations of the amounts. Pot the standard deviations
versus the amounts. Take the s5ecified standard deviation at the
corres5onding L" amount from the 5ot.
. S5ecificity with rea sam5es
Use sam5es with anaytes. Check
5eak 5urity with a diode-array detector and/ or a mass seective
detector. Run the sam5e under different chromatogra5hic coumns
and/or with different coumns.
9. Ruggedness Check 5recision and accuracy in
different a-oratories
. Ro-ustness
Systematicay change chromatogra5hic
conditions (eam5es: coumn tem5erature,fow rate, gradient
com5osition, 5 of mo-ie 5hase, detector waveength). Check
infuence of 5arameters on se5aration and/or 5eak areas.
Possi-e Parameters for ethod Vaidation
S5ecificity
Seectivity
Precision
Re5eata-iity
ntermediate 5recision
Re5roduci-iity
ccuracy
Trueness
ias
Linearity
Range
Limit of detection
Limit of quantitation
Ro-ustness
Ruggedness
SELECTIVITYAND SPECIFICITY
The terms 80l0ctivitv and 8p0ci1icitv are oIten used
interchangeably.
The term 8p0ci1ic generally reIers to a method that
produces a response Ior a single analyte only while
the term 80l0ctiv0 reIers to a method that provides
responses Ior a number oI chemical entities that may
or may not be distinguished Irom each other. II the
response is distinguished Irom all other responses, the
method is said to be selective. Since there are very
Iew methods that respond to only one analyte, the
term 80l0ctivitv is usually more appropriate.
The USP monograph deIines selectivity oI an analytical
method as its ability to measure accurately an analyte in the
presence oI interIerence, such as synthetic precursors,
excipients, enantiomers, and known (or likely) degradation
products that may be expected to be present in the sample
matrix. Selectivity in liquid chromatography is obtained by
choosing optimal columns and setting chromatographic
conditions, such as mobile phase composition, column
temperature, and detector wavelength.
igure1 Eam5es of 5ure and im5ure PLC 5eaks. The chromatogra5hic
signa does not indicate any im5urity in either 5eak. S5ectra evauation,
however, identifies the 5eak on the eft as im5ure.
PRECISION AND REPRODUCIBILITY
The precision oI a method is the extent to which the
individual test results oI multiple iniections oI a series
oI standards agree.
The measured standard deviation can be subdivided
into three categories:
repeatability
intermediate precision
reproducibility
Repeatability is obtained when the analysis is
carried out in one laboratory by one operator
using one piece oI equipment over a relatively
short time span. At least
Five or six determinations oI
Three diIIerent matrices at
Two or three diIIerent concentrations
should be done and the relative standard
deviation calculated.
The acceptance criteria Ior precision depend very
much on the type oI analysis.
While Ior compound analysis in pharmaceutical
quality a control precision oI better than RSD is
easily achieved, Ior biological samples the precision
is more like 5 at the concentration limits and 0
at other concentration levels.
Intermediate precision is a term that has been deIined
by ICH as the long-term variability oI the
measurement process and is determined by
comparing the results oI a method run within a single
laboratory over a number oI weeks.
A method`s intermediate precision may reIlect
discrepancies in results obtained by diIIerent
operators, Irom diIIerent instruments, with standards
and reagents Irom diIIerent suppliers, with columns
Irom diIIerent batches, or by a combination oI these.
The o-ective of intermediate 5recision
vaidation is to verify that in the same
a-oratory the method wi 5rovide the
same resuts once the deveo5ment 5hase
is over.
Reproducibility as deIined by ICH represents the
precision obtained between laboratories (Table 5).
The obiective is to veriIy that the method will provide
the same results in diIIerent laboratories.
The reproducibility oI an analytical method is
determined by analyzing aliquots Irom homogeneous
lots in diIIerent laboratories with diIIerent analysts
and by using operational and environmental
conditions that may diIIer Irom but are still within the
speciIied parameters oI the method (interlaboratory
tests). Validation oI reproducibility is important iI the
method will be used in diIIerent laboratories.
Typical Variations Affecting a Method`s Reproducibility
--------------------------------------------------------------------------------------------------
DiIIerences in room temperature and humidity
Operators with diIIerent experience and thoroughness
Equipment with diIIerent characteristics (e.g., delay volume oI
an HPC system)
Variations in material and instrument conditions (e.g., in HPC,
mobile phases composition,
pH, Ilow rate oI mobile phase)
Equipment and consumables oI diIIerent ages
Columns Irom diIIerent suppliers or diIIerent batches
Solvents, reagents, and other materials with diIIerent quality
ACCURACY AND RECOVERY
The accuracy oI an analytical method is the extent to
which test results generated by the method and the
true value agree. The true value Ior accuracy
assessment can be obtained in several ways.
One alternative is to compare the results oI the
method with results Irom an established reIerence
method.
This approach assumes that the uncertainty oI the
reIerence method is known. Second, accuracy can be
assessed by analyzing a sample with known
concentrations (e.g., a certiIied reIerence material)
and comparing the measured value with the true value
as supplied with the material.
LINEARITY AND CALIBRATIONCURVE
The linearity oI an analytical method is its ability to
elicit test results that are directly, or by means oI
well-deIined mathematical transIormation,
proportional to the concentration oI analytes in
samples within a given range.
inearity is determined by a series oI three to six
iniections oI Iive or more standards whose
concentrations span 8020 oI the expected
concentration range.
The response should be directly or by means oI a
well-deIined mathematical calculation proportional to
the concentrations oI the analytes.
A linear regression equation applied to the results
should have an intercept not signiIicantly diIIerent
Irom zero.
Frequently the linearity is evaluated graphically in addition or
alternatively to mathematical evaluation.
The evaluation is made by visual inspection oI a plot oI signal
height or a peak area as a Iunction oI analyte concentration.
Because deviations Irom linearity are sometimes diIIicult to
detect two additional graphical procedures can be used.
The Iirst one is to plot the deviations Irom the regression line
versus the concentration or versus the logarithm oI the
concentration iI the concentration range covers several
decades.
For linear ranges the deviations should be equally distributed
between positive and negative values.
Another approach is to divide signal data by their respective
concentrations yielding the relative responses.
A graph is plotted with the relative responses on the Y axis
and the corresponding concentrations on the X axis on a log
scale.
The obtained line should be horizontal over the Iull linear
range.
At higher concentrations there will typically be a negative
deviation Irom linearity.
Parallel horizontal lines are drawn in the graph corresponding
to, Ior example, 95and 05oI the horizontal line.
The method is linear up to the point at which the plotted
relative response line intersects the 95line.
Figure 2 Graphical presentations oI linearity plot oI a caIIeine sample using HPC.
Plotting the sensitivity (response/amount) gives a clear indication oI the linear range.
Plotting the amount on a logarithmic scale has a signiIicant advantage Ior wide linear
ranges. Rc line oI constant response.
RANGE
The range oI an analytical method is the interval between the
upper and lower
levels (including these levels) that have been demonstrated to
be determined
with precision, accuracy, and linearity using the method as
written. The range is normally expressed in the same units as
the test results (e.g., percentage, ppm)
obtained by the analytical method.
LIMIT OF DETECTION AND QUANTITATION
The limit oI detection is the point at which a measured value is
larger than the uncertainty associated with it.
It is the lowest concentration oI analyte in a sample that can be
detected but not necessarily quantiIied.
In chromatography the detection limit is the iniected amount that
results in a peak with a height at least twice or three times as
high as the baseline noise level.
The limit oI quantitation is the minimum iniected
amount that gives precise measurements, in
chromatography typically requiring peak heights
0 to 20 times higher than baseline noise.
ROBUSTNESS
Robustness tests examine the eIIect operational parameters
have on the analysis results.
For the determination oI a method`s robustness a number oI
chromatographic parameters (e.g., Ilow rate, column
temperature, iniection volume, detection wavelength, or
mobile phase composition) are varied within a realistic range
and the quantitative inIluence oI the variables is determined. II
the inIluence oI the parameter is within a previously speciIied
tolerance the parameter is said to be within the method`s
robustness range.
For example, to compensate Ior column perIormance over
time.
%ank you