The application of fully automated on-line solid phase extraction in bioanalysis.

Area of Science:

• Analytical Chemistry
• Bioanalysis
• Chromatography

Background:

• Bioanalysis often requires robust sample preparation techniques.
• Solid phase extraction (SPE) and liquid-liquid extraction (LLE) are common off-line methods.
• Automation can improve efficiency and reproducibility in bioanalytical assays.

Purpose of the Study:

• To evaluate the performance of fully automated on-line solid phase extraction (SPE) for bioanalysis.
• To compare the on-line SPE assay performance with traditional off-line methodologies.
• To assess the impact of compound physicochemical properties and matrix variations on assay performance.

Main Methods:

• Application of fully automated on-line SPE using the Symbiosis platform.
• Development and optimization of off-line SPE and LLE methods.
• Evaluation of linearity, carryover, imprecision, and inaccuracy for both on-line and off-line methods.
• Investigation of ionization and selectivity modifications using different plasma batches.

Main Results:

• On-line and off-line methods demonstrated comparable performance in terms of linearity, carryover, imprecision, and inaccuracy.
• The automated on-line SPE approach was successfully applied to three compounds with diverse physicochemical properties.
• Assay performance remained consistent despite modifications in ionization and selectivity against different plasma batches.

Conclusions:

• Fully automated on-line SPE is a viable and comparable alternative to off-line methods for bioanalysis.
• The on-line methodology offers significant advantages for high-throughput analysis by reducing manual operations.
• This automated approach enhances efficiency and reproducibility in bioanalytical workflows.