Low Adsorption HPLC Columns Based on MaxPeak High Performance Surfaces

Importance of the Topic

High-performance liquid chromatography (HPLC) applications involving metal-sensitive analytes often suffer from poor peak shape, low recovery, and quantitative errors due to Lewis acid–base interactions between electron-rich compounds and metal surfaces in column hardware. The development of low-adsorption surfaces is critical for ensuring robust, accurate separations and reliable mass spectrometry data across fields such as pharmaceutical analysis, biotherapeutics, environmental monitoring, and metabolomics.

Objectives and Overview

This white paper presents the design, characterization, and performance evaluation of MaxPeak High Performance Surfaces (HPS) applied to HPLC column hardware (ACQUITY PREMIER columns) and sample vials/plates (QuanRecovery). The goals are to mitigate undesired adsorption of metal-sensitive compounds, maintain ultrahigh-pressure capabilities, and validate stability under demanding conditions.

Methods and Instrumentation

• Column hardware modified with a hybrid organic/inorganic MaxPeak HPS layer (ethylene-bridged siloxane network).
• Test columns: ACQUITY UPLC PREMIER and standard columns packed with BEH C18, CSH C18, or CSH Phenyl Hexyl stationary phases (1.7 µm, 130 Å, 2.1 × 50 mm).
• Mobile phases: aqueous buffers (hexylammonium acetate, ammonium formate) with organic modifiers (acetonitrile), acid or base stress conditions (TFA pH 1, NaOH pH 12).
• Detection by UV absorbance (260 nm, 246 nm) and MS (electrospray, positive/negative mode).
• Quality control: ATP recovery tests on column-free analytical setup; contact angle measurements for hydrophobicity.

Main Results and Discussion

• Oligonucleotide separations showed consistent peak heights across injections on PREMIER columns, whereas standard columns required extensive conditioning.
• Peptide analysis: PREMIER hardware reduced tailing and revealed minor deamidated variants (PENNYK peptide) with improved quantitation.
• LC-MS of citric acid cycle metabolites demonstrated elimination of metal adducts and enhanced signal quality.
• Stress testing: MaxPeak HPS frits retained >80% ATP recovery after 16 h at pH 1–12 and 60–90 °C, indicating broad chemical stability.
• Mass load dependency: PREMIER columns delivered up to 83% higher peak area at 2 ng hydrocortisone phosphate and maintained linear response across 2–200 ng loads, outperforming standard hardware.

Contributions and Practical Applications

The MaxPeak HPS approach enables analysts to:

• Reduce or eliminate chelators in mobile phases, preserving chromatographic selectivity and MS sensitivity.
• Minimize conditioning time and improve throughput for metal-sensitive assays.
• Achieve sharper peaks, higher recovery, and consistent quantitation at low analyte loads.
• Extend applicability to organic acids, organophosphates, oligonucleotides, peptides, glycans, and phospholipids.

Future Trends and Applications

Advances in surface engineering are poised to further enhance inertness and customization for specific chromatographic modes. Integration with automated sample preparation and adoption in high-throughput LC-MS workflows will expand applications in proteomics, lipidomics, and small-molecule quantitation. Ongoing development of hybrid materials may yield surfaces with tunable hydrophobicity, charge properties, and selective binding sites.

Conclusion

MaxPeak High Performance Surfaces on PREMIER columns and QuanRecovery consumables address the longstanding challenge of analyte adsorption on metal hardware without sacrificing UPLC performance. This technology delivers reproducible, high-quality chromatographic and MS results across a wide pH and temperature range, simplifying method development and improving robustness for metal-sensitive analyses.

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