5. System Suitability
• Evaluate the System Suitability parameter as per
the test method
• The result should be within the limit specified in
the test method
5
6. Method Precision
6
• 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.
7. Method Precision
7
• 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.
8. Method Precision
8
• 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%
9. 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
9
10. Linearity
• Ability of an method
to elicit a direct and
proportional response
to changes in analyte
concentration.
10
11. 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
11
13. Accuracy
13
• 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.
14. Robustness
14
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)
15. Robustness
15
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.
16. Robustness
16
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.
17. Ruggedness
17
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)
18. Ruggedness
18
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.
19. Ruggedness
19
Methodology :
Perform the bench top and refrigerator stability of the standard and test solution
for 24hrs and 48 hrs ,calculate the results each time against freshly prepared
standard 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.
