Theoretical repeatability assessment without repetitive measurements in gradient high-performance liquid...

Area of Science:

• Analytical Chemistry
• Chromatography
• Spectroscopy

Background:

• Evaluating measurement repeatability in High-Performance Liquid Chromatography with UV detection (HPLC-UV) traditionally requires numerous sample analyses.
• Existing methods for assessing standard deviation (SD) and detection limits are resource-intensive.

Purpose of the Study:

• To propose a time and material-saving method for evaluating HPLC area measurement repeatability.
• To demonstrate the theoretical calculation of measurement standard deviation (SD) and detection limits using Function of Mutual Information (FUMI) theory.
• To validate the proposed method against traditional repetitive measurements.

Main Methods:

• Application of Function of Mutual Information (FUMI) theory to analyze baseline noise stochastic properties.
• Theoretical evaluation of standard deviation (SD) and detection limits.
• Optimizing the number of noise data points for theoretical evaluation (512 data points identified).
• Incorporation of sample injection variability (relative SD) into the theoretical model.

Main Results:

• The FUMI theory-based method theoretically provides measurement standard deviation (SD) and detection limits.
• Optimal noise data points for evaluation determined to be 512.
• Theoretical area measurement RSDs closely matched those from repetitive measurements (n=6) across a wide concentration range.
• SD estimates from FUMI theory (n=1) showed high reliability, comparable to 31 repetitive measurements.

Conclusions:

• The FUMI theory offers a viable, resource-efficient alternative for assessing HPLC measurement repeatability.
• The method accurately predicts measurement performance without repetitive sample analysis.
• This approach significantly reduces the time and materials needed for method validation in HPLC-UV analysis.