Dr Pramod Kumar Pandey, BSc (Hons), MSc, PhD, is the Founder of PharmaGuru.co and an Analytical Research Expert with 31+ years of experience in the pharmaceutical industry. He is recognized for driving analytical innovation, research excellence, and regulatory-compliant quality advancements across leading Indian and global pharmaceutical companies. He can be contacted at admin@pharmaguru.co
earn the key differences between analytical method development and validation using chromatographic, titrimetric, and mass spectrometric methods with case studies and FAQs.
Difference Between Analytical Method Development and Validation: HPLC, GC, LCMS, GCMS, Titrimetric & Spectroscopic Methods with Case Studies and FAQs
Both Analytical Method Development and Validation play a vital role in pharmaceutical development in managing quality, safety and efficacy. Analytical Method Development involves designing a reliable technique to identify and quantify chemical substances, while Analytical Method Validation confirms that this developed method consistently delivers accurate, precise, and reproducible results for its intended purpose.
In this blog, I will share key differences between Analytical Method Development and Validation with multiple case studies and FAQS.
Analytical Method Development and Validation
What is Analytical Method Development?
Analytical Method Development is the process of creating a reliable technique to identify and quantify chemical substances. It’s typically the first step in the lifecycle of an analytical procedure.
Purpose:
To design an analytical method that is suitable for the intended purpose.
Key Activities:
Selection of method type (e.g., HPLC, UV-Vis, GC, titration, FTIR, LCMS)
Optimisation of parameters (e.g., mobile phase, detection wavelength, temperature)
Assessing specificity, sensitivity, and reproducibility in early stages
Establishing a rough protocol to proceed toward validation
Example:
A pharmaceutical company is developing a new oral drug containing a novel API (Active Pharmaceutical Ingredient). Scientists begin method development to quantify this API using HPLC.
They choose a C18 column
Test different mobile phases (e.g., phosphate buffer with acetonitrile)
Determine the best detection wavelength (e.g., 245 nm)
Optimise the flow rate for peak resolution
What is Analytical Method Validation?
Analytical Method Validation is the process of proving that the developed method consistently produces reliable and accurate results for its intended use.
Purpose:
To demonstrate that the method is fit for regulatory and operational use, usually under ICH guidelines.
Optimisation of Conditions: It contains procedures for optimising the method.
STP or Standard Test Procedure: It includes detailed procedures for performing pharmaceutical analysis.
Method development report or method verification report: Parameters like Specificity, Precision, Linearity, DL and QL are performed, and the report is prepared to establish the reliability of the method
Elements of Analytical Method Validation
The following are the main elements of the Analytical method development:
Approved Method or Monograph:It is required to prepare an analytical method validation protocol.
Analytical Method validation protocol: It is required to perform an analytical method validation study.
Analytical Method validation report: It contains the validation study work.
Method validation parameters: Depending upon the criticality of the method all or some of the following parameters can be performed during method validation:
Specificity: The specificity parameter is used to establish that there is no interference between an adjacent impurity peak or an impurity peak with the main peak or a blank peak with the main peak or an impurity peak.
Accuracy and Recovery: The closeness of the measured value to the true value (the reference or accepted standard).
Precision: The degree of reproducibility of results under normal operating conditions, typically evaluated by assessing repeatability and intermediate precision.
Linearity: The ability of the method to give results that are directly proportional to the concentration of analyte over a specified range.
Range: The interval between the upper and lower concentration limits within which the method can provide accurate and precise results.
DL or Detection Limit: The smallest amount of analyte that can be reliably detected but not necessarily quantified.
QL or Quantification Limit: The lowest concentration of analyte that can be reliably quantified with acceptable precision and accuracy.
Robustness: The method’s capacity to remain unaffected by small variations in operational conditions
Stability of Solutions: The Stability of Solutions is performed to avoid any failure during analysis in sequence
What are the Quality Attributes of Analytical Method Development and Validation?
The following main quality attributes play a vital role in Analytical Method Development and Validation:
Responsibilities
GLP (Good laboratory practice)
GMP (Good Manufacturing Practice)
SOP (Standard Operating Procedure)
Data integrity
CAPA (corrective and preventive action)
OOS (out of specification)
OOT (out of trend)
Technology transfer (TT)
Regulatory compliance
Audit management &
QBD (Quality by design) approach
Who is responsible for analytical method development and validation?
The following are the responsibilities of different professionals/ departments:
Analytical Professional/Validation Specialists: They are responsible for performing analytical method development and validation studies
Quality Control (QC) Professional: QC professionals are involved in method validation in reproducibility tests. They are not involved in Analytical method development
Regulatory Affairs Personnel: They coordinate with various regulatory agencies such as FDA, EMA and provide answers to any queries related to AMD or AMV.
What is the role of SOP, GLP, GMP, data integrity, CAPA, OOS, OOT and OOS in analytical method development and validation?
Method development and method validation intrinsically linked to GLP (Good Laboratory Practice), GMP (Good Manufacturing Practice), Data Integrity, SOP, CAPA (Corrective and Preventive Action), OOS (Out of Specification), OOT (Out of Trend) It has happened.
GLP (Good Laboratory Practice):GLP ensures that laboratory processes, including method development, are properly documented, validated, and performed with high standards of quality and integrity. GLP governs analytical research and chemical research activities.
GMP (Good Manufacturing Practice):GMP guidelines require that the methods used in the analysis of pharmaceuticals be thoroughly validated.
Data Integrity: Method development and validation activities must comply with stringent data integrity principles (e.g., ALCOA-Attributable, Legible, Contemporaneous, Original, and Accurate). This ensures that the data generated during these processes is accurate, complete, and traceable.
CAPA (Corrective and Preventive Actions): During method development and validation, any observed issues or deviations must be investigated and addressed using CAPA. If a method does not perform as expected or shows inconsistencies, CAPA procedures are implemented to correct the problem and prevent future occurrences.
OOS (Out of Specification): OOS refers to results that fall outside predefined acceptance criteria. It is only related to validation and not to development.
How are analytical method development and validation helpful in technology transfer?
Analytical method development and validation provide the technical basis to ensure that transferred technology produces consistently high-quality products, complies with regulatory standards, and minimizes the risks associated with transfer between sites or teams.
The analytical method development and validation are helpful in technology transfer in the following ways:
Ensuring Consistency and Reproducibility:
Method Development: Developing robust analytical methods ensures that the same results can be obtained regardless of where the testing is performed. This is especially important when transferring technology across different sites or between different teams.
Validation: The validation of these methods demonstrates that they consistently produce accurate results within specified limits, ensuring that the quality of the product is maintained even when produced in new locations or scaled-up processes.
Regulatory Compliance:
Regulatory agencies (such as the FDA, EMA, etc.) require validated analytical methods for product testing, quality control, and stability studies. Proper validation ensures that the methods comply with these regulations, facilitating smooth technology transfer by reducing the risk of regulatory issues during inspections or audits.
Risk Mitigation:
Analytical methods provide critical data on the quality attributes of the product, such as purity, assay, degradation products, and impurities. During technology transfer, having validated methods ensures that the new production process or manufacturing site will maintain product quality, thus minimising the risks of batch failures or product recalls.
Reproducibility Across Different Stages of Production:
Analytical method development and validation ensure that these methods are capable of maintaining the quality at different production stages
Data Integrity
Validated analytical methods ensure the integrity of the data generated throughout the technology transfer process.
Training and Knowledge Transfer
The analytical methods that have been developed and validated are often critical tools for training personnel at the receiving site. By transferring both the method itself and the rationale behind its validation, organisations can ensure that the new team has the necessary knowledge to maintain consistent product quality.
How are analytical method development and validation helpful for the Drug master file?
The analytical method development and validation play a vital role in the submission, approval, and ongoing compliance of pharmaceuticals such as:
Ensures Consistency and Quality Control: It controls the quality of the pharmaceuticals such as assay, purity, impurity profile, and stability.
Supports Regulatory Compliance: Regulatory authorities require that all analytical methods used in the testing of pharmaceuticals must be validated according to the approved protocol
Demonstrates the Drug’s Safety and Efficacy: It plays a vital role in maintaining safety and efficacy of the pharmaceuticals.
Enables Stability Testing: Validated stability indicating methods (SIMs) are used to test the stability of pharmaceuticals over time, ensuring that they maintain their efficacy and safety during storage. Stability studies, supported by SIMs, are required for regulatory submissions.
Improves Traceability and Documentation: A well-documented validation process provides a clear, reproducible method for analysing the drug. This documentation is crucial for the DMF, as it offers regulatory agencies detailed evidence that the product can consistently meet quality standards.
Facilitates Communication with Regulatory Authorities: The inclusion of comprehensive method development and validation data helps in addressing regulatory questions or concerns during the review process, providing a clear understanding of how the drug is tested and ensuring all regulatory requirements are met.
What is the role of analytical method development and validation in audit inspection?
Analytical verification plays an important role in audit inspection in the pharmaceutical industries. Method validation review is required in almost all audits.
How do Analytical Method Development and Validation fulfil Regulatory Requirements?
Analytical Method Development and Validation play a vital role in DMF filing and answering the deficiency letter queries.
How is the Quality by Design (QBD) approach helpful in analytical method development and validation?
The QBD approach helps analytical method development and validation by providing a scientific rationale in the understanding of critical parameters of the method, reducing variability, and ensuring reliability and compliance. This structured RFT (right first time) approach not only improves the efficiency of method development and validation but also reduces risk.
The QbD approach helps in analytical method development and validation in the following ways:
Understanding the Analytical Target Profile (ATP): It outlines the purpose of the method and the critical parameters that must be controlled to achieve the desired results. This focus helps avoid trial-and-error during method development and guides the optimisation of method parameters.
Identification of Critical Quality Attributes (CQAs): In QbD, the Critical Quality Attributes (CQAs) are identified early and ensures that the method delivers reliable and consistent results under various conditions.
Risk Assessment and Control of Variables: It is very helpful in identifying the critical variables which may lead to failure of AMV or AMD.
Design of Experiments (DoE): QbD helps in using the design of experiments (DOE) to systematically explore the effect of various factors on method performance. DOE helps in identifying the optimal combination of conditions (e.g., temperature, pH, mobile phase composition) for reliable method development. It is also helpful in finalising the validation protocol and in performing robustness tests.
Analytical Method Transfer: QbD helps in Analytical method transfer from the Analytical laboratory to QC (quality control).
Cost-effectiveness: QbD helps in reducing Analytical method development and validation cost
Regulatory Compliance: QbD provides a more robust rationale for decisions made during method development, allowing for more streamlined regulatory submissions, approvals, and compliance
What is Difference between Analytical Method Development and Validation?
The following are the main differences between Analytical Method Development and Validation
Elements
AMD
AMV
Definition
AMD is an innovative work and it is performed to establish analytical conditions to perform analysis
AMV is a skill-based work and it is performed to establish the reliability of the developed analytical method
Selection of appropriate techniques
Required (e.g., HPLC, GC, spectrometry)
Not required. Established technique is used
Optimizing conditions
Required(e.g., mobile phase, temperature, detector settings)
Not required.
STP/Monograph
Not needed
Either STP or Monograph is required to perform AMV
Protocol
Not needed
Approved protocol is needed to perform validation
Literature report
Very helpful for AMD
Not needed
Samples
AMD may be performed using pure or impure samples
Representative samples with valid COAs are required to perform AMV
Standards/Impurities
Only selected parameters are used to find out the criticality of the method and avoid any surprise/ failure during complete validation
Standards/Impurities (as mentioned in the monograph) with valid COAs are required to perform AMV
Validation parameters
Only selected parameters are performed to find out the criticality of the method and avoid any surprise/ failure during complete validation
All parameters (as defined in the validation protocol) are performed to find out the criticality of the method and avoid any surprise/ failure during complete validation
Report
A complete validation report is prepared
Performed under GLP and GMP environments
Approval process
Analytical project coordinator and Analytical head
Analytical/Validation project coordinator, Analytical /Validation head, QC head and QA head
GLP/GMP requirement
Performed under GLP environment
AMD may be performed using an individual sample or contaminated /impure sample or standard
Data integrity
Must be maintained
Must be maintained
Guidelines
Not essential
Required
SOP
Not essential
Required
LIR
Not required
Required
QBD
Play a vital role in designing the development experiment
Play a vital role in designing the validation protocol
How helpful are Analytical Method Development and Validation?
Analytical Method Development and Validation are very helpful in:
Ensuring Product Quality, Safety and Efficacy:
Analytical Method Development: Method must be able to provide specific, accurate, precise and reproducible results
Validation: Validation ensures that the analytical method is suitable for the intended purpose
Supporting Pharmaceutical Development
During Pharmaceutical development, analytical methods are used to characterize, identify and quantify the pharmaceuticals
Regulatory Compliance
Regulatory agencies such as the FDA and EMA require that pharmaceutical products meet quality standards. Analytical methods must be developed and validated to comply with regulatory guidelines..
Validation provides documented evidence that the method is reliable. If the method is not validated as per the guidelines, then many deficiency letter-related questions may arise.
Ensuring Consistency
Analytical methods ensure consistency in production batches during QC analysis
Controlling Impurities in Pharmaceuticals
Be it purchasing the ingredients, reaction mix, intermediates and the final API method is used at every stage to monitor the quality
Stability Studies
Stability indicating method plays a vital role about formation of degradation impurities during stability studies
Batch Release and Quality Control
In quality control (QC), analytical methods are used to test and release or reject raw materials, in-process samples, and APIs. Hence, the method must be reliable and validated to avoid any failure
Risk Mitigation
Analytical method validation helps to minimise analytical method risks and ensure the reliability of results.
What are the Advantages and Disadvantages of Analytical Method Development and Validation?
The following are the advantages of the Analytical method development and Validation:
Method reliability
Regulatory Compliance
Consistency in analytical results
Maintaining pharmaceutical quality
Cost Efficiency
Risk Management
Data Integrity and Traceability
Reproducibility Across Laboratories
The following are the disadvantages of the Analytical method development and Validation:
Both Analytical method development and Validation take a lot of time
Needs knowledge as well as expertise
Costly activities
Conclusion
While Method Development is about creativity, optimisation, and scientific exploration, Method Validation is about discipline, documentation, and regulatory compliance. Both are essential for ensuring product quality and patient safety.
Organisations like ICH, FDA, EMA, and USP provide regulatory guidelines (e.g., ICH Q2(R2)).
What is the difference between method development and validation?
Analytical Method Development involves designing a reliable technique to identify and quantify chemical substances, while Analytical Method Validation confirms that this developed method consistently delivers accurate, precise, and reproducible results for its intended purpose.
