Thermo Scientific HyperSep Columns Application Notebook – Issue 1

Importance of the Topic

Solid-phase extraction (SPE) is a cornerstone of modern analytical chemistry, providing efficient cleanup, concentration and fractionation of diverse sample matrices before chromatographic or mass-spectrometric analysis. Advances in SPE technology, such as the Thermo Scientific HyperSep column family, address challenges in pharmaceuticals, environmental monitoring, forensic toxicology and food safety by reducing matrix interferences, improving sensitivity and streamlining workflows.

Study Goals and Overview

This application notebook presents an extensive overview of HyperSep SPE chemistries, guidelines for phase selection and detailed protocols for hundreds of target analytes across biological, environmental, forensic and food matrices. It aims to remove uncertainty in sample preparation by applying systematic SPE science, offering:

• Phase selection criteria based on analyte properties (polarity, pKa, matrix complexity)
• Optimized conditioning, loading, washing and elution steps
• Demonstrated recoveries for key compounds (drugs, pesticides, toxins)

Methodology and Instrumentation

Method development follows a six-step SPE procedure:

• Sample pretreatment (filtration, pH adjustment, dilution)
• Column conditioning (organic solvent and aqueous equilibration)
• Sample loading at controlled flow rate
• Selective washing to remove interferences
• Targeted elution to recover analytes
• Drying and reconstitution for analysis

SPE phases include polymeric reversed-phase (Retain PEP, Retain-CX/AX), silica reversed-phase (C18, C8, Phenyl), normal-phase (Silica, Florisil, Cyano, Diol), hypercrosslinked carbon (Hypercarb) and mixed-mode ion exchangers (Verify-CX/AX, SAX, SCX). Instrumentation encompasses GC/MS, LC/UV, HPLC, LC-MS/MS and GC-MS platforms, illustrating broad compatibility.

Main Findings and Discussion

Wash and elution profiles for benzodiazepines, beta-agonists and environmental pollutants highlight the need for analyte-specific optimization. For example, Retain-CX columns achieved >90 % recovery of basic drugs at wash strengths below breakthrough thresholds, while Retain PEP efficiently captured moderately polar herbicides with minimal co-extraction of humic substances. Mixed-mode sorbents simplified multi-class drug screens in biological fluids, reducing solvent usage and processing time compared to liquid–liquid extraction.

Benefits and Practical Applications

Key advantages of HyperSep SPE workflows include:

• Enhanced selectivity and reduced matrix effects for LC and GC analysis
• Lower solvent consumption compared to traditional extraction methods
• Scalability for manual or automated high-throughput applications
• Wide pH compatibility and stable recoveries across analyte classes

Applications range from acetaminophen quantitation in serum, beta-blocker screening in urine, pesticide residue analysis in water, to multi-mycotoxin extraction in cereal grains.

Future Trends and Opportunities

Emerging directions in SPE technology will focus on:

• Miniaturized, high-throughput formats for micro-SPE and 96-well plates
• Novel sorbent chemistries targeting polar emerging contaminants (per- and polyfluoroalkyl substances, polar metabolites)
• Integrated on-line SPE–LC-MS systems for real-time monitoring
• Green solvents and reusable cartridges to improve sustainability
• AI-driven method development for rapid phase and protocol selection

Conclusion

Thermo Scientific HyperSep columns provide a versatile SPE toolkit that addresses the evolving demands of analytical laboratories. By combining tailored sorbent chemistries with optimized protocols, users can achieve consistent, high-recovery extractions across diverse matrices and analytes. The systematic approach outlined in this notebook reduces method development time and enhances data quality for both routine and cutting-edge analyses.

References

No specific literature references were listed in the source document.