In this slide contains Analytical method validation as per USP Presented by: P.SUDHEER KUMAR (Department of pharmaceutical analysis).RIPER, anantapur

1. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
ANALYTICAL METHOD
VALIDATION AS PER USP
1
Presented by
P. Sudheer Kumar
(20L81S0714)
Department of Pharmaceutical
Analysis

2. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Contents
• Introduction
• Objective
• Types of analytical procedures to be validated
• Validation parameters as per USP
• Conclusion
• References
2

3. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Introduction
• Validation is the documented act of proving that any procedure,
process, equipment, material, activity or system actually leads to
the expected result.
• Analytical method validation is a process of documenting/proving
that an analytical method provides analytical data acceptable for
the intended use.
• A pharmaceutical drug product must meet all its specifications
through out its shelf-life.
3

4. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
• The method of analysis used must be validated. This is required to
ensure the product’s safety and efficacy through out all phases of
its shelf-life
Objective:
• The main objective of analytical validation is to ensure that a
selected analytical procedure will give reproducible and reliable
results that are adequate for the intended purpose.
• This is applicable to all the procedures either pharmacopoeial and
nonpharmacopoeial.
4
Cont…

5. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
*The required validation parameters also termed “ analytical
performance characteristics”, depends upon the type of analytical
method. Pharmaceutical analytical methods are characterized into 5
types.
 Identification tests
 Potency assays
 Limit tests for the control of impurities
 Impurity tests- quantitative
 Specific tests
5
Types of analytical procedures to be
validated

6. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
1. Specificity
2. Linearity
3. Range
4. Accuracy
5. Precision
6. Limit of detection
7. Limit of quantitation
8. Robustness
9. SST
6
Validation parameters as per USP

7. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
1. Specificity :
Specificity is the ability to assess unequivocally the analyte in
presence of components which may be expected to be present.
Determination :
• Identification tests
• Assay and impurity test(s)
 Impurities are available
 Impurities are not available
7

8. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
2. Linearity:
• The Ability of the method to obtain test results that are directly
proportional to concentration within a given range. Method:
dilution of stock solution/separate weightings
• Minimum 5 concentrations are used.
8

9. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
3.Range Definition:
The interval between the upper and lower concentrations of analyte
in the sample that have been demonstrate to have a suitable level of
precision, accuracy, and linearity.
• Established by confirming that the method provides acceptable
degree of linearity, accuracy, and precision.
• Specific range dependent upon intended application of the
procedure.
9

10. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cont…
• Assay: 80 to 120% of test concentration.
• Content uniformity: 70 to 130% of test concentration.
• Dissolution: 20% to 120%
• Impurities reporting level: 120% of specification limit (with
respect to test concentration of API)
10

11. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
4. Accuracy Definition:
The accuracy of an analytical procedure is the closeness of agreement
between the values that are accepted either as conventional true
values or an accepted reference value and the value found.
Determination of
 Assay
i. Drug substance
ii. Drug product
 Impurities (quantitation)
11

12. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
• Recommended data: Assessed by 9 determinations over a
minimum of 3 concentration levels covering a specified range.
• Limit:
i. Typical accuracy of the recovery of the drug substance is
expected to be about 99 – 101%.
ii. Typical accuracy of the recovery of the drug product is expected
to be about 98 – 102%.
12
Cont…

13. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
5. Precision Definition:
The closeness of agreement (degree of scatter) between a series of
measurements obtained from multiple samplings of the same
homogeneous sample.
• Precision includes:
i. Repeatability
ii. Intermediate Precision
iii. Reproducibility
13

14. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
i. Repeatability:
• Repeatability expresses the precision under the same operating
conditions over a short interval of time.
• Repeatability should be assessed using a minimum of 9
determinations covering the specified range.
ii. Intermediate precision
• Intermediate precision expresses variations within laboratories,
such as different days, different analysts, different equipment etc.
14

15. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 15
iii. Reproducibility:
Reproducibility expresses the precision between laboratories.

16. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
6. Limit of detection:
It is the lowest amount of analyte in a sample which can be detected
but not necessarily quantitated.
7. Limit of quantitation:
It is the lowest amount of analyte in a sample which can be
quantitatively determined with suitable precision and accuracy.
16

17. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Determination of LOD and LOQ
Limit of detection:
• Based on visual examination.
• Based on standard deviation of response and slope.
• Signal to noise ratio 2:1 or 3:1
Limit of quantitation:
• Based on visual examination.
• Based on standard deviation of response and slope.
• Signal to noise ratio 10:1
17

18. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
8. Ruggedness Definition:
The ruggedness of an analytical method is the degree of
reproducibility of test results obtained by the analysis of the same
samples under a variety of conditions, such as different laboratories,
different analysts, different instruments, different days, etc.
i Certain may include
i. Source
ii. Concentration and stability of solution
iii. Heating rate
iv. Column temperature
v. Humidity
18

19. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Robustness Definition:
“The robustness of an analytical procedure is a measure of its
capacity to remain unaffected by small, but deliberate variations in
method parameters and provides an indication of its reliability
during normal usage”. Determination: The evaluation of robustness
should be considered during the development phase and depends
on the type of procedure under study.
19

20. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Cont…
Variations may include:
• stability of analytical solution
• variation of pH in a mobile phase
• different column (lot/supplier)
• temperature
• flow rate
20

21. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
9. System suitability
• System suitability testing is an integral part of many analytical
procedures.
• The tests are based on the concept that the equipment,
electronics, analytical operations and samples to be analyzed
constitute an integral system that can be evaluated as such.
• System suitability testing has been recommended by USP in
HPLC procedures.
21

22. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
Conclusion
• When the method is properly validated consistent, reliable and
accurate results are obtained.
• Analytical method validation is an important analytical tool to
ensure the accuracy and specificity of the analytical procedures
with a precise agreement.
• Validation of analytical methods is also required by regulations.
• Hence it is very important to validate any analytical method that
has been developed.
22

23. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
References
1. Gupta P. C method validation of analytical procedures. pharmatutor..
2. Lopez P, Buffoni E, Pereira F, Vilchez Quero J. Analytical Method Validation.
Wide Spectra of Quality Control.
3. [Internet]. Who.int. 2016 [cited 2 March 2019].
4. https://www.who.int/medicines/areas/quality_safety/quality_assurance/Gu
ideline_Validation _AnalyticalMethodValidationQAS16-671.pdf
5. Daksh S. VALIDATION OF ANALYTICAL METHODS – STRATEGIES &
SINGFICANCE. International Journal of Research and Development in
Pharmacy and Life Sciences.
6. [Internet]. Ich.org. [cited 2 March 2019].
https://www.ich.org/fileadmin/PublicWebSite/ICH_Products/Guidelines/Qu
ality/Q2 _R1/Step4/Q2_R1Guideline.pdf
7. Sharma P. validation in pharmaceutical industry. 2nd ed. Delhi: Vandana
publications.
23

24. RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 24