Analytical Method Analytical Method Validation Validation

for for DissolutionDissolution

Bhanu Prakash. N

(Analytical R&D)

Email : bhanu.analytical@gmail.com

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Why Method Validation is Important

The purpose of analytical validation is to get consistent, reliable and

accurate data.

Definition of validation

• The objective of validation of an analytical procedure is to

demonstrate that it is suitable for its intended purpose.

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ICH/USP Validation Requirements

• Precision

• Specificity

• Linearity

• Accuracy

• Robustness

• Ruggedness

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System Suitability• Evaluate the System Suitability parameter as per

the test method• The result should be within the limit specified in

the test method

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Method Precision

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• The closeness of agreement (degree of scatter)

between a series of measurements obtained from

multiple samplings of the same homogeneous sample.

• Should be investigated using homogeneous, authentic

samples.

Method Precision

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• Perform the dissolution as per the test method and

calculate the % dissolution , average % dissolution, %

RSD and confidence interval.

• For multiple strength, conduct method precision for

highest, and lowest strength.

Method Precision

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• The method is consider ‘precise’, if the dissolution result

are NLT 95% of label amount and the % RSD of six units

should not less than 5.0%

Specificity-Placebo Interference

• Perform the dissolution as per the test method on weight of placebo in duplicate, equivalent to the amount present in tablets powder or capsules. In case of capsule take filled capsule with placebo

• The chromatogram of placebo should not show any peak at retention time of Active peak

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Linearity

• Ability of an method

to elicit a direct and

proportional response

to changes in analyte

concentration.

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Linearity

• Demonstrate the linearity in the range of 10% to 150% of the target concentration.

• Prepare series of standard solution (not less than 5 are recommended ) using drug substance

• Plot the graph of peak area versus actual concentration(ug per mL)

• The correlation coefficient should not less than 0.999

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Accuracy

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Closeness of the test results obtained by the method to the true value.

Accuracy

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• Prepare the test solution(s) in triplicate of each spike

level and analyze as per the test method. Calculate the

% recovery.

• The method is considered ‘Accurate’ if the average

recovery is NLT 95.0% of label amount.

Robustness

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Robustness : Definition: It is a measure of method's ability to remain unaffected by small but

deliberate variations in method parameters.

The following are the typical method parameters need to change deliberately and verify during method validation :

Flow rate (+/- 0.2ml/minutes) Mobile phase composition (+/- 10% of organic phase) Column oven temperature (+/- 5°C) pH of buffer in mobile phase (+/- 0.2 units) Filter suitability (At least two filters)

Robustness

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Methodology :

a) Mobile phase variation: Prepare the mobile phases by changing organic phase to +/-10% of the mobile phase composition.

b) Flow rate : Change the flow rate by +/- 0.2 ml/minutes of the target flow rate mentioned in test method.

c) Temperature of the Column : Change the temperature the column by +/- 5.0°C of the target temperature mentioned in Test method.

d) pH of the buffer of mobile phase : Prepare the mobile phases by changing the pH of buffer by +/- 0.2 units of the pH mentioned in the test method.

Robustness

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Methodology :

E) Filter Suitability : Prepare the test solution as per test method and filter through

two different types filters. Analyse the sample as per test method and compare

the results against unfiltered / centrifuged sample.

Acceptance Criteria :

For Variations : 1. System suitability should meet the acceptance criteria as per test method.

For filter suitability : Difference between the % dissolution should not be more than 3.0% against the unfiltered / centrifuged sample.

Ruggedness

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Definition:

It is a measure of method reproducibility under variable conditions within specified test parameters of test method.

The following are the typical method parameters need to tested during method validation :

Analyst-to-Analyst variability. Column-to-Column variability. System-to-System variability. Different days. Different Laboratories. Stability of Solutions and mobile phase. (At least for 48 hours)

Ruggedness

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Methodology :

• Perform the precision as mentioned in precision of test method by using different systems, different columns, different analyst , in different Laboratories and on different day.

Ruggedness

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Methodology :

Perform the bench top and refrigerator stability of the standard and test solutionfor 24hrs and 48 hrs ,calculate the results each time against freshly preparedstandard solution.If 24hrs stability is not meeting the acceptance criteria,perform the stability studies on hourly basis on bench top and in refrigerator.

Acceptance Criteria :

The dissolution % Difference should not be more than 3.0% from initial to 1 and 2 day .

Similarity factor of standard should be NLT 0.98 and NMT 1.02.

Thank Thank YouYou

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