Bioanalytical method validation – EMEA guidelines MCQs With Answer

Introduction: Bioanalytical method validation – EMEA guidelines MCQs With Answer is designed to help M.Pharm students master key principles of bioanalytical method validation as outlined by the European Medicines Agency. This set focuses on practical regulatory expectations: accuracy, precision, calibration curve requirements, selectivity, matrix effect, recovery, stability studies, incurred sample reanalysis, dilution integrity, carryover, and batch acceptance criteria. Questions are tailored to reinforce both conceptual understanding and application to laboratory practice, emphasizing acceptance limits and method performance parameters. Use these MCQs to prepare for exams and practical lab assessments, and to ensure compliance with robust validation standards required for bioanalytical assays.

Q1. Which acceptance criterion for accuracy does the EMEA guideline recommend for quality control (QC) samples, excluding LLOQ?

• Within ±10% of nominal
• Within ±15% of nominal
• Within ±20% of nominal
• Within ±25% of nominal

Correct Answer: Within ±15% of nominal

Q2. According to EMEA, what is the acceptable precision (expressed as CV) for the lower limit of quantification (LLOQ)?

• CV ≤10%
• CV ≤15%
• CV ≤20%
• CV ≤25%

Correct Answer: CV ≤20%

Q3. How many non-zero calibration standards does the EMEA guideline generally recommend to construct a reliable calibration curve?

• At least 4
• At least 6
• At least 8
• At least 10

Correct Answer: At least 6

Q4. For back-calculated concentrations of calibration standards, what acceptance limit is recommended by EMEA (except for LLOQ)?

• Within ±10% of nominal
• Within ±15% of nominal
• Within ±20% of nominal
• Within ±25% of nominal

Correct Answer: Within ±15% of nominal

Q5. Which of the following best defines dilution integrity as per EMEA expectations?

• The ability to accurately measure diluted blank samples
• The ability to accurately measure samples diluted into the calibration range without loss of accuracy and precision
• The ability to dilute calibration standards only
• The ability to dilute internal standard concentrations

Correct Answer: The ability to accurately measure samples diluted into the calibration range without loss of accuracy and precision

Q6. What is the minimum acceptable percentage of incurred sample reanalysis (ISR) pairs that should be within ±20% for reanalysis according to EMEA guidance best practice?

• 50%
• 60%
• 67%
• 75%

Correct Answer: 67%

Q7. Which stability study assesses the effect of multiple freeze–thaw cycles on analyte integrity in the matrix?

• Bench-top stability
• Long-term stability
• Autosampler stability
• Freeze–thaw stability

Correct Answer: Freeze–thaw stability

Q8. In method validation, what does selectivity primarily assess under EMEA guidance?

• Linearity of response across concentrations
• Ability to separate analyte from endogenous matrix components and co-medications
• Instrument calibration accuracy
• Internal standard stability

Correct Answer: Ability to separate analyte from endogenous matrix components and co-medications

Q9. What is the recommended acceptance criterion for accuracy and precision at LLOQ according to EMEA?

• Both within ±10% and CV ≤10%
• Both within ±15% and CV ≤15%
• Both within ±20% and CV ≤20%
• No specific criteria for LLOQ

Correct Answer: Both within ±20% and CV ≤20%

Q10. Which practice is recommended to evaluate matrix effects for LC–MS/MS assays in EMEA guidance?

• Comparison of peak areas from neat solutions to post-extraction spiked samples across multiple lots of matrix
• Only running calibration standards in solvent
• Measuring internal standard in water
• Using a single lot of matrix for all tests

Correct Answer: Comparison of peak areas from neat solutions to post-extraction spiked samples across multiple lots of matrix

Q11. What is the purpose of quality control (QC) samples during validation and routine analysis per EMEA?

• To serve as replacement calibration standards
• To monitor assay performance for accuracy and precision within runs and between runs
• To calibrate the instrument daily
• To test only the internal standard response

Correct Answer: To monitor assay performance for accuracy and precision within runs and between runs

Q12. Which of the following is an EMEA-recommended practice for carryover assessment?

• Inject a blank after the upper limit of quantification (ULOQ) and ensure no significant analyte signal
• Only check carryover at the beginning of the run
• Rely on instrument vendor specifications without testing
• Inject only low concentration samples to assess carryover

Correct Answer: Inject a blank after the upper limit of quantification (ULOQ) and ensure no significant analyte signal

Q13. Recovery assessment in bioanalytical method validation primarily determines what?

• Absolute signal intensity of calibration standards
• Efficiency and reproducibility of the extraction procedure for the analyte from the matrix
• Stability of analyte in stock solution
• Linearity of the detector response

Correct Answer: Efficiency and reproducibility of the extraction procedure for the analyte from the matrix

Q14. According to EMEA, how many independent validation runs are generally recommended for method validation accuracy and precision assessment?

• One run with multiple replicates
• Two runs on the same day
• At least three separate validation runs
• Five runs across different instruments only

Correct Answer: At least three separate validation runs

Q15. What is the primary role of an internal standard in quantitative bioanalytical assays as per EMEA guidance?

• To serve as an extra calibration point
• To correct for sample preparation variability, matrix effects, and instrument fluctuation
• To increase analyte concentration
• To act as a preservative in the sample

Correct Answer: To correct for sample preparation variability, matrix effects, and instrument fluctuation

Q16. What does autosampler stability evaluate in method validation?

• Stability of analyte when stored frozen for years
• Stability of processed samples kept in the autosampler at controlled temperature over the typical run time
• Stability of stock solutions at room temperature
• Stability of calibrators after light exposure

Correct Answer: Stability of processed samples kept in the autosampler at controlled temperature over the typical run time

Q17. Which statement about calibration curve weighting is consistent with EMEA recommendations?

• No weighting should ever be used for calibration curves
• Weighting (e.g., 1/x or 1/x2) can be applied to improve fit, especially over wide concentration ranges
• Only 1/x2 is acceptable for all assays
• Weighting replaces the need for QC samples

Correct Answer: Weighting (e.g., 1/x or 1/x2) can be applied to improve fit, especially over wide concentration ranges

Q18. For long-term stability studies, which condition should be demonstrated according to EMEA guidance?

• Analyte stability at intended storage temperature for the maximum anticipated storage time of study samples
• Only short-term bench stability for 1 hour
• Stability only in solvent, not in matrix
• Stability over 10 years without testing

Correct Answer: Analyte stability at intended storage temperature for the maximum anticipated storage time of study samples

Q19. What is the EMEA-recommended acceptance criterion for the percentage deviation of QC samples during routine batch analysis?

• All QCs must be exactly at nominal
• At least 67% of QCs per concentration should be within ±15% (±20% for LLOQ)
• Only one QC needs to be within ±25%
• 50% of QCs must be within ±10%

Correct Answer: At least 67% of QCs per concentration should be within ±15% (±20% for LLOQ)

Q20. Which action is appropriate if significant matrix effect is observed across multiple lots during validation?

• Ignore it if calibration fits well
• Investigate and modify sample preparation, chromatographic separation, or use a suitable internal standard to compensate
• Change the analyte structure
• Reduce the number of QC samples

Correct Answer: Investigate and modify sample preparation, chromatographic separation, or use a suitable internal standard to compensate

Download