Recovery Studies in Peptide Analysis
Recovery measures how much analyte is detected after sample preparation compared with the amount expected.
Why recovery matters
Peptides can be lost through:
- incomplete dissolution
- adsorption
- filtration
- precipitation
- degradation
- extraction inefficiency
- transfer losses
Spike-recovery design
Known analyte is added to the matrix at multiple levels. The measured increase is compared with the added amount.
Pre- and post-process spikes
A pre-process spike evaluates the entire preparation. A post-process spike can help isolate matrix effects or instrument response.
Acceptance criteria
Criteria should reflect intended use, concentration, and method capability.
Frequently asked questions
Can 100% recovery prove no matrix effect?
No. Matrix effects may still alter response in more complex ways.
Why test multiple levels?
Loss may be concentration-dependent.
Can recovery exceed 100%?
Yes, due to bias, interference, or calibration error.
Should recovery use the same containers as routine testing?
Yes. Surface effects are part of the process.
Key takeaways
Recovery studies reveal hidden sample-preparation losses and matrix bias. They should reproduce the complete routine workflow.
References
- ICH Q2(R2). Validation of Analytical Procedures.
- ICH Q14. Analytical Procedure Development.
- FDA. Analytical Procedures and Methods Validation for Drugs and Biologics.
TSMS Labs educational disclaimer: For laboratory research and educational purposes only. Not for human consumption. This content is not medical, clinical, or regulatory advice.