KIM3471

VALIDATION – METHOD VALIDATION

VALIDATION
Validation is a word of Latin origin and means confirmation. Independent
testing and measurement procedures to demonstrate that the
performance of a device, method, or system conforms to specified
conditions.
The definition of validation in the ISO 17025 standard is "objectively
testing and documenting the suitability of a method or measurement
procedure for specified purposes." made in the form.

Validation ; always produces the same quality and under the same
conditions, ensuring that the quality of all manufactured products is the
same.
Why Validation is done? VALIDATION TYPES

• As required by GMP rules, 1.BUILDING/INFRASTRUCTURE

• To ensure quality, to minimize variability 2. MACHINERY/EQUIPMENT
• To create well-controlled reliable processes, 3. PROCESS VALIDATION
• To have better knowledge of equipment and 4. ANALYTICAL METHOD VALIDATION
processes, 5. CLEANING VALIDATION
• To reduce cost, 6. COMPUTER SYSTEM VALIDATION
• To increase productivity,
•To increase coordination, communication and
information flow.
Analytical Method Validation
Analytical method validation is to prove the validity of the method used in analysis.
Apart from this definition, the definition of validation may differ according to the
guidelines. According to USP Supplement 9; It is to show whether the performance
characteristics of the method are suitable for the purpose of the targeted analytical
applications, by laboratory studies. The ICH Guideline Q2A states that the main
purpose of validation is to demonstrate the fitness of an analytical procedure for its
intended purpose. There may be differences in the guidelines as the purpose of
interest may be different.

The purpose of analytical method validation is to:

*Identify and identify sources of potential errors *Determine

whether the method is fit for purpose
*Prove that the method is suitable for making predictions
*Fulfill the requirements of the GMP or FDA.
When method validation is performed:
* When a standard method is applied for the first time in a laboratory
* When a new method is developed for an analysis
* When a change is made in the method used
* When a validated method will be used in another laboratory or when it will
be used with a different person or a different device
*when it is understood that there is a change in the performance of the
method over time
Method validation; It can be confused with qualification and verification.
Pre-clinical studies, phase 1 and phase 2 studies in drug development have not been
fully validated, but qualified methods can be used. Qualified methods do not have
fully validated performance test documents. But it can evaluate the variability and
reliability of control methods.
Verification, on the other hand, is the confirmation, under laboratory conditions, of a
method (pharmacopeia method, etc.) for which performance criteria have been
determined through interlaboratory studies.

The level of validation of the methods varies according to the established

performance criteria. Accordingly:
*Verification is sufficient if a method has been validated by an interlaboratory study.
*A method validated by an interlaboratory study; however, if a sample with a different
structure is to be studied, accuracy, precision, detection limit and detection limit
should be confirmed by adding a new matrix to the verification.
*If a method has been validated by an intralaboratory study rather than an
interlaboratory study, validation parameters should be added to the verification
parameters as much as possible.
*If a method has been published in a scientific journal and certain performance
parameters have been studied, validation parameters should be added to the
verification parameters as much as possible.
*A method has been published in a scientific journal; however, if no performance
parameters have been examined, full internal validation should be performed.
*If a method does not have any performance criteria or is developed in-house, full
internal validation should be performed.
Method validation; It is part of the entire
validation process, consisting of at least four
independent steps, such as software
validation, device validation and
qualification, analytical method validation,
and system compliance. The entire
validation process starts with validated
software and validated qualified system.
Then the method is developed and validated
using a qualified system. They should then
be checked together to confirm expected
performance limits. This process is called
system compatibility testing. Finally, the
accuracy of sample analysis data collected
from such a system is confirmed by
appropriate validation controls.
Analytical Method Validation According to Types of Methods
USP and ICH divide analytical methods into four categories. These are:
Category 1: Analyzes for the quantification of main components or active
additives
Category 2: Determination of impurity or decomposition products
Category 3:Determination of performance characteristics
Category 4: Identification (Identification) tests
Analytical performance characters used according to method types
 General Method Validation Steps
Method validation consists of at least five different steps. These are: system
qualifications, sampling, sample preparation, analysis and data evaluation.
System Qualifications
The overall assessment of system qualifications is that the instrument is suitable
for the desired analysis, all material such as reagents, reference substances,
internal and external standards are suitable for use in analytical determination,
analysts have appropriate qualifications and training, and historical documentation
such as analytical procedure is considered predetermined. It is done to verify that it
has been approved according to the criteria. If the general system qualifications are
ignored, any problem will become larger and it will be very difficult to identify the
source of the problem.
Sampling
Sampling step assists in the process of selecting the representative fraction of the
material to be used in the study. The selection of the appropriate sampling method
is of great importance. Because it assures that the correct representative sample
is selected with a meaningful statistical result.

Sample Preparation
Sample preparation is a key element of successful method validation. Sample
preparation represents 60-80% of operational efficiency and operating cost in the
analytical laboratory. The literature on sample preparation is abundant and well
documented. However, the investigator should note that the choice of the
particular preparation procedure depends on the analytes, their analytical
concentration, sample matrix, sample size, and instrument technique.
Analysis
An instrumentation analysis method is used to obtain qualitative and quantitative
information from samples with an acceptable level of uncertainty. Selection of
specific analyzes; The chemical properties of the analytical species, the amount in
the sample, the matrix of the sample, speed and cost are taken into account.

Data Evaluation
The purpose of data evaluation is to summarize results and derive
meanings/inferences from partial datasets using mathematical and statistical
approaches. Data evaluation provides useful data selection and results plots for
inputs and outputs and is crucial in the validation process.
Validation Plan
Planning should be done before starting the validation/verification work. The plan should
be prepared by considering factors such as laboratory facilities, analytical feasibility and
customer demands, and should be able to answer the following questions.
•What is the purpose of using the method?
•Is a qualitative or quantitative result required?
• In what chemical form does the analyte exist? (free, bound, different chemical
compounds)
•Is the analyte regionally dispersed?
• What is the area where the method is applied?
•Is there interference from the sample structure or other analytes?
How much is the sample amount and is it homogeneous?
•Will sampling and subsampling be done?
• In what interval is the method close to the detection limit or at a higher level?
•Are there legal limits for the analyte?
•How will accuracy be ensured?
• Will the results be compared with other laboratory results?
•What are the necessary environmental conditions?
•Is there a resource constraint? What are the financial limits?
The validation/verification plan should include the parameters, time schedule and details
to be evaluated in the study.
Method Validation in Pharmaceutical Laboratories

Analytical methods determine the quality of drugs, their effects on living and stability.
When choosing analysis method, the following situations are taken into account.
•Is the description of the drug in the product correct?
•What is the percentage of active ingredient in the formula?
•Does the formula contain additional impurities?
• At what rate is the active substance released in the formulation and at what rate is itby the body absorbed?
•Does the identity and purity of the pure drug substance to be used meet the specification?
• What are the concentrations of certain impurities in the pure drug substance?
•What is the concentration of the drug substance in tissues and biological fluids?
•What are the pKa values, distribution coefficients, any drug substance during development?
Chemical analysis is used to answer these questions. The quality of any product required the quality of
may deviate from the standard, but any analysis has to be will meet the required standard precise as it.
Pharmaceutical researchers in each country have different way with their own conditions and
purposes for the development of quality methods.
To standardize this situation, the analytical quality control of methods has been determined by
ICH.
Validation according to ICH requiring;
•Identification experiments
•Quantitative experiments for impurities
•Quantitative experiments to determine limits for control of impurities
• Quantitative analysis of active and other ingredients
•Assays used to determine storage, storage conditions and stability of drugs.
Types of method validation
1- Analytical method validation: procures used to prove the suitability of the analytical
method
2- Bioanalytical validation: They are procedures that prove the appropriateness of the
analytical method to be used in the evaluation of human samples.
3- Bioanalytical method validation: It both as test samples and as assay reagents
refers to biologically based substances used.
4-Method feature validation: method feature with more than initially expected
Acceptance of the limits (for example, ±20% of accuracy), preclinical development for
example, the limits (for example, ±10%)
Validation Parameters
There are various internationally accepted validation criteria. Criteria determined by institutions
such as ISO/IEC, ICH, US EPA, US FDA, USP, cGMP are used.
Validation parameters describe the parameters to be validated from the analysis methods.
Validation Parameters
There are various internationally accepted validation criteria. Criteria determined by institutions
such as ISO/IEC, ICH, US EPA, US FDA, USP, cGMP are used.
Validation parameters describe the parameters to be validated from the analysis methods.