ISOLUTE® HYDRO DME+ Dual Mode Extraction 96-well Plates and Columns

Importance of the Topic

Urine analysis by LC-MS/MS frequently encounters matrix effects from pigments, salts, urea, creatinine and residual enzymes. Robust cleanup is essential to maintain sensitivity, reproducibility and instrument uptime. ISOLUTE HYDRO DME+ provides a streamlined solution that enhances assay performance and minimizes downtime.

Objectives and Study Overview

This whitepaper evaluates the performance of Biotage ISOLUTE HYDRO DME+ 96-well plates and 400 mg/3 mL columns for urine sample preparation prior to LC-MS/MS analysis. Key aims include assessing matrix removal efficiency, hydrolysis effectiveness and signal stability compared to a conventional dilute-and-shoot approach.

Methodology and Instrumentation

• Sample Workflow: In-well enzymatic hydrolysis, addition of acetonitrile, mixing and pass-through cleanup under low vacuum or positive pressure.
• Dual Mode Extraction: Combines liquid partitioning with sorbent scavenging to remove interferents while retaining target analytes.
• In-Situ Hydrolysis: HYDRO frit technology enables enzymatic reaction at elevated temperature directly in the plate or column.
• Instrumentation: LC-MS/MS system; Biotage ISOLUTE HYDRO DME+ plates (Part 970-0400-PZ01) and columns (Part 970-0040-BZ); vacuum or positive pressure manifold.

Main Results and Discussion

• Matrix Removal: Effective clearance of urinary pigments, salts, urea, creatinine and enzyme residues, yielding visually clear extracts.
• Signal Stability: Maintained consistent analyte response for over 800 injections, compared to a >50% sensitivity drop after 400 injections with dilute-and-shoot.
• Hydrolysis Efficiency: Equivalent to traditional offline protocols, facilitating direct transfer of existing methods.
• Operational Robustness: Extended intervals between source cleans reduce instrument downtime and maintenance costs.

Benefits and Practical Applications of the Method

• Enhanced Sensitivity: Prolonged stable signal benefits toxicology, clinical and pharmacokinetic assays.
• High Throughput: Eliminates evaporation and transfer steps, supporting automated workflows.
• Cost Reduction: Decreases solvent consumption, labor and maintenance requirements.

Future Trends and Applications

Dual mode extraction is poised to expand into other biological matrices such as plasma and saliva. Ongoing sorbent innovations may enable multiplexed residue analyses and improved selectivity for emerging biomarkers. Miniaturization and green chemistry approaches will further enhance sustainability and throughput.

Conclusion

ISOLUTE HYDRO DME+ plates and columns offer a fast, automatable and effective cleanup strategy for urine LC-MS/MS workflows. By combining dual mode extraction with in-situ hydrolysis, the technology delivers superior matrix removal, prolonged signal stability and seamless integration with existing protocols.

References

No literature references were provided in the source document.