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Checklist for Analytical Method Validation

Analytical Method Validation is to be performed for new analysis methods or for current methods when any changes are made to the procedure, composition of the drug product and synthesis of the drugs substances. Common types of analytical procedures that can be validated.


What is meant by method validation?
According to the FDA, “Analytical method validation is the process of demonstrating that an analytical procedure is suitable for its intended purpose. The methodology and objective of the analytical procedures should be clearly defined and understood before initiating validation studies.

What is the purpose of method validation?
Method validation data provide information that enables the comparability of results from samples analyzed in different laboratories and using different methods to be assessed. Method validation is an essential part of the process of ensuring that measurement results reported to customers are correct.

What is method validation in the laboratory?
Method validation is a demonstration of the method's suitability by determining the accuracy of the test results as well as the uncertainty and traceability of measurements. Method validation is needed for proving whether the new method is fit for the purpose or specified samples.

What are the types of validation?
Prospective validation
Retrospective validation.
Concurrent validation.
Revalidation.
 
What are LOD and LOQ in method validation?
The limit of detection (LOD) and limit of quantification (LOQ) are two important performance characteristics in method validation. LOD and LOQ are terms used to describe the smallest concentration of an analyte that can be reliably measured by an analytical procedure.


What is the difference between method validation and verification?
In conclusion, method validation is usually applied to an “in-house method” developed by a laboratory; while method verification is applied to a “compendia method or previously validated method” when it is being used in a particular laboratory for the first time.

What is the difference between method validation and Process Validation?
The major difference in the process and product validation is their methodologies. Process validation is aimed at ensuring different inputs will guarantee a similar end product while product validation aims to ensure the end product works effectively to its intended purpose; meeting user needs and requirements.

What are the parameters of validation?
The Validation parameters are accuracy, precision, linearity and application range, limit of detection (LOD), limit of quantitation (LOQ), selectivity/specificity, recovery and robustness/ruggedness.

Why is analytical method validation important?
Validation of analytical test methods is important for every stage of the drug development life cycle. Methods must be shown to be acceptable for their intended use and must provide accurate and reliable results at every stage of development, from pre-clinical trials through approved and marketed products.


How do you validate a test method?
The accuracy of an analytical procedure is defined as how close the test results of the parameters are for a specific analyte compared to the true measure of these parameters.
Precision.
Specificity.
Limit of detection.
Limit of quantitation.
Linearity.
Range.
Robustness.


How is the LOD value calculated?
LOD may also be calculated based on the standard deviation of the response (Sy) of the curve and the slope of the calibration curve (S) at levels approximating the LOD according to the formula: LOD = 3.3(Sy/S).

What is the blank limit?
Limit of blank (also known as the critical value) is the highest quantity value that is likely to be observed, with a stated probability, for a blank material. Related concepts. Limit of detection (LoD) Related tasks. Estimating the detection limit of a measurement system.


What is the limit of linearity?
Limit of linearity (LOL). The concentration at which the calibration curve departs from linearity. The extends from the LOQ to LOL is called the dynamic range. A well-defined relationship between detector response and analyte concentration is crucial for quantitative analysis.

Checklist for Analytical Method Validation (Chemical)

Assay/Related Substances


TEST

ASSAY/RELATED SUBSTANCES

PARAMETER

No.

DOCUMENTS REQUIRED

AVAILABILITY

 

 

 

 

 

 

 

 

 

 

Specificity

1

Testing Method

 

2

Acceptance criteria

 

3

Chromatogram/spectrum for following solutions:

 

 

Standard

 

Sample

 

Blank/Placebo

 

Spike solution

 

System suitability tests

 

4

Impurities available

 

 

Peak purity for PDA detector or spike solution for non-PDA detector

 

Relative Retention Time (RRT)

 

Resolution (R) where applicable

 

5

Impurities not available

 

 

Stress data (minimum data for humidity, heat and light)

 

Peak purity

 

Relative Retention Time (RRT)

 

Resolution (R) where applicable

 

 

 

Linearity

1

Testing Method

 

2

Acceptance criteria

 

3

Minimum five (5) levels of standard solutions

 

4

Data for linear regression equation, Y-intercept, slope, r2 and linearity graph

 



 

Range

1

For the assay of drug substances: 80% - 120% of working

concentration (WC)

 

2

For the determination of an impurity: from the reporting level of an impurity to 120% of the specification

 

 

 

 

 

Accuracy

1

Testing Method

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range

 

4

Result: 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 (Repeatability)

1

Testing Method (using sample/product as the test solution)

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range, OR minimum six (6) replicates at 100% of the WC

 

4

Result: SD, RSD and confidence Interval

 

 

 

 

Precision (intermediate precision/ ruggedness)

1

Testing Method (using sample/product as the test solution)

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range, OR

minimum six (6) replicates at 100% of the WC

 

4

Cover at least 2 parameters among variation of analyst, date and equipment

 

5

Result: SD, RSD and confidence Interval

 




Detection Limit

1

Testing Method: visual observation / signal-to-noise / standard deviation of the response and the slope

 

2

If based on standard deviation of the response and the slope method

 

 

Minimum five (5) levels of standard solutions

 

Peak area values for all concentrations

 

Data for linear regression equation, Y-intercept, slope, r2 and linearity graph.

 

3

Calculation/formulation where applicable

 

4

Related Chromatogram(s) at LOD

 

5

Value of detection limit

 

Quantitation Limit

1

Testing Method: visual observation / signal-to-noise / standard deviation of the response and the slope

 

2

if based on visual observation method, accuracy and precision data at LOQ must be provided

 

3

If based on calibration curve method

 

 

Minimum five (5) levels of standard solutions

 

Peak area values for all concentrations

 

Data for linear regression equation, Y-intercept, slope, r2 and linearity graph.

 

5

Calculation/formulation where applicable

 

6

Value of quantitation limit

 

System

Suitability Testing

1

RSD, tailing factor, theoretical plate

 

2

Resolution (if two or more components)

 

Robustness (not mandatory)

1

Testing Method

 

2

Acceptance criteria

 

3

Result: refer acceptance criteria for accuracy and precision (repeatability)

 



Checklist for Analytical Method Validation (Chemical)

Disolution


TEST

DISSOLUTION

PARAMETER

No.

DOCUMENTS REQUIRED

AVAILABILITY

 

 

 

 

Specificity

1

Testing Method

 

2

Acceptance criteria

 

3

Chromatogram/spectrum for following solutions:

 

 

Standard

 

Sample

 

Blank/Placebo

 

Spike solution

 

System suitability tests

 

 

 

Linearity

1

Testing Method

 

2

Acceptance criteria

 

3

Minimum five (5) levels of standard solutions

 

4

Data for linear regression equation, Y-intercept, slope, r2 and linearity graph.

 

 

 

 

Range

1

Dissolution testing: ± 20% over the specified range

Example 1: if the specification is NLT 75% (Q) of the labelled amount is dissolved in 45 minutes, the validated range would be 60 – 100% of the label claim

Example 2: if the specification for a controlled released product covers a region from 20% after 1 hour, up to 90%, after 24 hours, the validated range would be 0 – 110% of the label claim

 

 

 

 

 

Accuracy

1

Testing Method

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range

 

4

Result : 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 (Repeatability)

1

Testing Method (using sample/product as the test solution)

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range, OR minimum six (6) replicates at 100% of the WC

 

4

Result: SD, RSD and confidence Interval

 

 

 

 

Precision (intermediate precision/ ruggedness)

1

Testing Method (using sample/product as the test solution)

 

2

Acceptance criteria

 

3

Minimum three (3) levels of concentration in triplicates covering the specified range , OR minimum six (6) replicates at 100% of the WC

 

4

Cover at least 2 parameters among variation of analyst, date and equipment

 

5

Result: SD, RSD and confidence Interval

 

System

Suitability Testing

1

RSD, tailing factor, theoretical plate

 

2

Resolution (if two or more components)

 

Robustness (Not Mandatory)

1

Testing Method

 

2

Acceptance criteria

 

3

Result: refer acceptance criteria for accuracy and precision (repeatability)

 


PEOPLE ALSO READ: Validation Master Plan


Commonly Acceptance Criteria

PARAMETER

ACCEPTANCE CRITERIA

Specificity

Absence of interfering peaks in the placebo, impurity demonstrate specificity

Pass peak purity test (particularly for related substances test)

Linearity

r2 ≥ 0.995

y-intercept at 100% working concentration ≤ 2%

Accuracy

Measured recovery within 95% - 105% or mean difference ± 2% & CI

Precision

(Repeatability)

RSD ≤ 2.0% & CI

Precision (intermediate precision/

ruggedness)

RSD ≤ 2.0% & CI

or

mean difference ± 2% & CI

 

Detection Limit

LOD peak must be visible

If based on standard deviation of the response and the slope method, DL = 3.3 Ó/S

If based on signal to noise, S/N= 3:1 or 2:1

 

 

Quantitation Limit

if based on visual observation method, accuracy and precision data at LOQ must be ± 20%

If based on standard deviation of the response and the slope method, DL = 10 Ó/S

If based on signal to noise, S/N= 3:1 or 2:1

 

System Suitability Testing

RSD ≤ 2%

Theoretical plate/column efficiency, N ≥ 2000

Tailing factor < 2

Resolution > 2

Robustness

(Not mandatory)

Refer acceptance criteria for accuracy and precision (repeatability)


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