Uncertainty in quantitative thin-layer chromatography

Area of Science:

• Analytical Chemistry
• Metrology

Background:

• Globalization necessitates enhanced control over analytical measurement variability.
• The error budget model, established by the Bureau International des Poids et Mesures, is a standard for measurement uncertainty but not directly applicable to separation techniques.
• Quantitative measurement uncertainty in separation techniques remains a challenge.

Purpose of the Study:

• To investigate the applicability of the error budget model as a planning tool for thin-layer chromatographic (TLC) method validation.
• To develop and test a computer program for quantifying uncertainty components in quantitative TLC.
• To assess whether the error budget approach can substitute traditional validation procedures.

Main Methods:

• Experimental validation of the error budget model using thin-layer chromatography (TLC).
• Development of a computer program to quantify uncertainty in quantitative TLC.
• Testing with TLC and high-performance TLC plates, various samples, manual and automated application, and multiple scanning modes (UV-vis, video documentation, remission, transmission, fluorescence).

Main Results:

• The error budget model can be utilized as a planning tool in the validation of quantitative TLC methods.
• A computer program successfully quantitated uncertainty components in quantitative TLC.
• Calculated uncertainty values using the error budget model were comparable to those from traditional validation procedures.

Conclusions:

• The error budget approach is a valuable planning tool for quantitative TLC method validation but cannot substitute the complete validation process.
• While it can predict critical points in quantitative TLC, it does not confirm the validity of a chosen chromatographic procedure.