How to Process ISOLUTE® SLE+ Plates and Columns with Biotage® PRESSURE+ 96 and 48 Positive Pressure Manifolds

Importance of the Topic

Solid Phase Extraction (SPE) and Supported Liquid Extraction (SLE) are foundational techniques in bioanalytical chemistry, offering rapid sample cleanup, high analyte recovery, and minimized matrix effects. Transitioning from conventional liquid–liquid extraction, the ISOLUTE SLE+ system paired with positive pressure manifolds addresses challenges such as emulsion formation, operator variability, and limited throughput.

Objectives and Overview

This application note describes a robust workflow for processing ISOLUTE SLE+ plates and columns in 96- and 48-well formats using Biotage PRESSURE+ positive pressure manifolds. It details setup requirements, stepwise sample loading, incubation, and elution procedures designed to ensure reproducible and high-quality extract preparation.

Methodology and Instrumentation

Key protocol steps:

• Place ISOLUTE SLE+ extraction plate and collection plate on the PRESSURE+ tray, ensuring proper alignment.
• Load aqueous samples (e.g., plasma, urine) by pipette into each well.
• Lower the manifold, apply 1–2 psi positive pressure to initiate sample passage, then pause for a 5-minute equilibration.
• Add elution solvent, reapply pressure for solvent flow into the collection plate, and repeat if multiple elutions are required.

Instrumentation details:

• Biotage PRESSURE+ 96 and 48 Positive Pressure Manifolds
• ISOLUTE SLE+ plates and columns (200 µL, 400 µL, 1 mL formats)
• Gas supply with adjustable flow control for precise pressure regulation

Main Results and Discussion

The PRESSURE+ system delivers highly uniform pressure across all wells, translating into consistent flow rates and analyte recoveries. ISOLUTE SLE+ plates, constructed with RENSA RP resin (90 µm particles, 120 Å pores, 400 m²/g surface area), demonstrate excellent chemical compatibility and mechanical stability. The protocol effectively processes both low- and high-viscosity samples without emulsion formation, achieving reproducible extraction efficiencies across batches.

Benefits and Practical Applications

Applications span pharmaceutical drug analysis, food additive quantification, amino acid profiling, and chemical intermediate monitoring. Major benefits include:

• Parallel processing of up to 96 samples reduces hands-on time and increases throughput.
• Elimination of emulsions and minimized solvent usage compared to traditional LLE.
• User-friendly setup with universal plate compatibility and built-in safety features.

Future Trends and Opportunities

Integration with automated liquid handling platforms and online monitoring systems will further streamline workflows. Emerging sorbent materials and miniaturized well formats are expected to extend applicability to ultra-low volume, high-sensitivity assays. Software-driven pressure control and data tracking may enhance traceability and compliance in regulated environments.

Conclusion

Combining ISOLUTE SLE+ technology with Biotage PRESSURE+ positive pressure manifolds offers a reliable, high-throughput solution for bioanalytical sample preparation. The standardized pressure-driven workflow ensures reproducible recoveries, operational safety, and compatibility with diverse sample matrices.

References

• Biotage Application Note AN747.V.1