Introduction: Analytical method validation is a fundamental part of pharmaceutical analysis for B. Pharm students, ensuring methods are reliable, accurate and fit for their intended purpose. Key validation parameters include accuracy, precision, specificity, linearity, range, limit of detection (LOD), limit of quantitation (LOQ), robustness and system suitability. Understanding ICH guidelines (such as Q2(R1)), QC acceptance criteria, calibration strategies and statistical evaluation helps in developing compliant methods for drug assay, impurity testing and stability studies. Mastery of these principles ensures consistent product quality, regulatory compliance and confident interpretation of analytical data. Now let’s test your knowledge with 30 MCQs on this topic.
Q1. What is the primary purpose of analytical method validation in pharmaceuticals?
- To check instrument ownership
- To ensure a method is suitable, reliable and reproducible for its intended use
- To replace method development
- To train analysts only
Correct Answer: To ensure a method is suitable, reliable and reproducible for its intended use
Q2. Which ICH guideline is most often cited for analytical method validation?
- ICH Q3C
- ICH Q8
- ICH Q2(R1)
- ICH M7
Correct Answer: ICH Q2(R1)
Q3. Which parameter assesses the closeness of agreement between the measured value and the true value?
- Precision
- Accuracy
- Robustness
- Specificity
Correct Answer: Accuracy
Q4. Repeatability, intermediate precision and reproducibility are subcategories of which validation parameter?
- Linearity
- Precision
- LOD/LOQ
- Range
Correct Answer: Precision
Q5. Which parameter demonstrates the method’s ability to measure the analyte response proportionally across a specified concentration range?
- Specificity
- Linearity
- System suitability
- Robustness
Correct Answer: Linearity
Q6. The lowest amount of analyte that can be detected but not necessarily quantified is called:
- Limit of Quantitation (LOQ)
- Limit of Detection (LOD)
- Range
- Accuracy
Correct Answer: Limit of Detection (LOD)
Q7. Which approach is commonly used to estimate LOD and LOQ based on the calibration curve?
- Signal-to-noise ratio only
- Standard deviation of the response and slope (3.3σ/s and 10σ/s)
- Visual inspection of chromatograms
- Use of internal standard concentration
Correct Answer: Standard deviation of the response and slope (3.3σ/s and 10σ/s)
Q8. Specificity (or selectivity) in method validation refers to:
- The method’s throughput
- The method’s ability to measure the analyte in presence of other components like impurities, excipients and degradation products
- The method’s cost-effectiveness
- The reproducibility between labs
Correct Answer: The method’s ability to measure the analyte in presence of other components like impurities, excipients and degradation products
Q9. Robustness testing evaluates:
- How the method responds to deliberate small variations in method parameters
- Interpersonal variability only
- The financial robustness of a lab
- The method’s ability to detect impurities at trace levels
Correct Answer: How the method responds to deliberate small variations in method parameters
Q10. System suitability tests are performed:
- Only after method validation is complete
- Before sample analysis to ensure the system is working properly
- Only for dissolution methods
- Only for microbiological methods
Correct Answer: Before sample analysis to ensure the system is working properly
Q11. Which statistical measure is commonly used to express precision as a relative value?
- Mean recovery
- Standard deviation
- Relative Standard Deviation (RSD or %RSD)
- Correlation coefficient (r)
Correct Answer: Relative Standard Deviation (RSD or %RSD)
Q12. In accuracy studies, which experimental design is often used for recovery testing?
- Placebo challenge
- Standard addition at multiple concentration levels (e.g., 80%, 100%, 120%)
- Only single concentration spike
- System suitability injections
Correct Answer: Standard addition at multiple concentration levels (e.g., 80%, 100%, 120%)
Q13. Which of the following indicates acceptable linearity for a calibration curve in many analytical methods?
- Correlation coefficient (r) ≤ 0.90
- Correlation coefficient (r) ≥ 0.999 always
- Correlation coefficient (r) typically ≥ 0.99 and residuals randomly distributed
- RSD of standards > 10%
Correct Answer: Correlation coefficient (r) typically ≥ 0.99 and residuals randomly distributed
Q14. Which validation parameter addresses potential matrix effects in complex samples (e.g., plasma)?
- Linearity
- Specificity/selectivity and recovery studies
- System suitability
- LOD estimation
Correct Answer: Specificity/selectivity and recovery studies
Q15. What is the usual acceptance criteria for percent recovery in accuracy experiments for assay methods?
- Recovery between 50–60%
- Recovery typically within 98–102% for assay, depending on method and concentration
- Recovery > 200%
- Recovery exactly 100% only
Correct Answer: Recovery typically within 98–102% for assay, depending on method and concentration
Q16. Method transfer between laboratories requires demonstration of:
- Only instrument brand matching
- Equivalence of method performance (precision, accuracy, specificity) between labs
- Different sample preparation
- No documentation
Correct Answer: Equivalence of method performance (precision, accuracy, specificity) between labs
Q17. Which parameter defines the upper and lower concentration limits where the method is validated with acceptable accuracy and precision?
- LOD and LOQ
- Range
- Specificity
- Robustness
Correct Answer: Range
Q18. Signal-to-noise ratio method for LOQ typically uses which S/N values?
- S/N of 1 for LOQ
- S/N of 3 for LOQ
- S/N of 10 for LOQ and 3 for LOD
- S/N of 100 for LOQ
Correct Answer: S/N of 10 for LOQ and 3 for LOD
Q19. Which practice helps demonstrate specificity for chromatographic methods?
- Running blank, placebo, standard, sample and forced degradation samples and checking peak purity
- Only measuring a single standard
- Using visual color comparison
- Skipping impurity checks
Correct Answer: Running blank, placebo, standard, sample and forced degradation samples and checking peak purity
Q20. Robustness experiments commonly include deliberate changes in:
- Ambient lighting conditions only
- Chromatographic parameters like flow rate, pH, column temperature, mobile phase composition
- Analyst salaries
- Supplier invoices
Correct Answer: Chromatographic parameters like flow rate, pH, column temperature, mobile phase composition
Q21. For method precision (repeatability), which is a typical experimental requirement?
- One replicate measurement
- Multiple injections or analyses (e.g., six independent preparations) at a single concentration
- Only system suitability samples
- Only external lab data
Correct Answer: Multiple injections or analyses (e.g., six independent preparations) at a single concentration
Q22. In forced degradation studies, the main goal is to:
- Produce market samples
- Generate degradation products to demonstrate method specificity and stability-indicating capability
- Reduce method sensitivity
- Increase sample throughput
Correct Answer: Generate degradation products to demonstrate method specificity and stability-indicating capability
Q23. Which calibration approach can correct for matrix effects and recovery losses in bioanalytical methods?
- External calibration only
- Standard addition or use of an internal standard
- Ignoring matrix effects
- Using a single-point calibration without QC
Correct Answer: Standard addition or use of an internal standard
Q24. Which of the following best describes ‘system suitability’ parameters for HPLC?
- System suitability refers to instrument warranty
- Parameters such as theoretical plates, tailing factor, resolution and %RSD of peak areas
- Only detector wavelength check
- Only mobile phase pH
Correct Answer: Parameters such as theoretical plates, tailing factor, resolution and %RSD of peak areas
Q25. When validating an impurity method, what is an important acceptance criterion?
- Impurities must not co-elute with the main peak and LOQ must be adequate to detect specification levels
- Impurities can co-elute with the main peak
- Only the main peak matters
- No need to report LOQ
Correct Answer: Impurities must not co-elute with the main peak and LOQ must be adequate to detect specification levels
Q26. Which statistical test is useful to compare precision between two analysts or labs?
- t-test for means
- F-test for variances
- Chi-square for proportions
- Kruskal-Wallis for medians only
Correct Answer: F-test for variances
Q27. In validation reports, which section documents how the method was proven to be stability-indicating?
- System suitability section only
- Specificity/forced degradation studies section
- Supply chain section
- Financial summary section
Correct Answer: Specificity/forced degradation studies section
Q28. Which is a common acceptance limit for %RSD in system suitability for assay methods?
- %RSD ≤ 20%
- %RSD ≤ 2% for replicate injections, depending on method
- %RSD ≥ 50%
- No limit is needed
Correct Answer: %RSD ≤ 2% for replicate injections, depending on method
Q29. What is the difference between verification and validation in analytical methods?
- Verification is confirming a validated method in a new lab; validation is establishing method performance originally
- They are identical terms with no difference
- Verification is done only for instruments
- Validation is optional
Correct Answer: Verification is confirming a validated method in a new lab; validation is establishing method performance originally
Q30. Which documentation is essential to include at the end of a method validation study?
- Validation protocol, raw data, calculations, statistical analysis, conclusions and acceptance criteria
- Only a one-line summary
- Only the final chromatogram
- Only the analyst’s resume
Correct Answer: Validation protocol, raw data, calculations, statistical analysis, conclusions and acceptance criteria
I am a Registered Pharmacist under the Pharmacy Act, 1948, and the founder of PharmacyFreak.com. I hold a Bachelor of Pharmacy degree from Rungta College of Pharmaceutical Science and Research. With a strong academic foundation and practical knowledge, I am committed to providing accurate, easy-to-understand content to support pharmacy students and professionals. My aim is to make complex pharmaceutical concepts accessible and useful for real-world application.
Mail- Sachin@pharmacyfreak.com