Introducing Atlantis BEH Z-HILIC: A Zwitterionic Stationary Phase Based on Hybrid Organic/Inorganic Particles

Importance of the Topic

Hydrophilic interaction chromatography (HILIC) is a key technique for separating polar compounds in fields such as metabolomics, pharmaceutical analysis and food testing. A stationary phase with strong retention, broad pH stability and minimal surface charge interactions is essential for method flexibility and reproducibility.

Objectives and Study Overview

This study introduces a new zwitterionic sulfobetaine stationary phase based on ethylene-bridged hybrid (BEH) particles. Key aims included evaluation of pH range (2–10), column efficiency, retention, selectivity, batch-to-batch reproducibility and performance with metal-sensitive analytes.

Methodology and Instrumentation

The performance of the BEH Z-HILIC phase was assessed through:

• Efficiency vs flow rate tests using acenaphthene (void) and cytosine (retained) markers.
• Retention and selectivity experiments with neutral and ionic test compounds.
• Batch-to-batch reproducibility studies across 17 production lots.
• Acid and base stability challenges at 70 °C over extended cycles.
• Analysis of metal-sensitive analytes (ATP, ADP, AMP) with MaxPeak HPS hardware.

Used Instrumentation

• ACQUITY UPLC I-Class, H-Class and Premier Systems
• ACQUITY UPLC PDA and TUV detectors
• Atlantis Premier BEH Z-HILIC, BEH Amide and BEH HILIC columns (1.7 µm, 2.1 × 50 mm)
• Empower 3 FR4 chromatography data software

Main Results and Discussion

• High efficiencies: Maximum 12 100 plates (H) for cytosine, plate height 4.1 µm.
• Strong retention: Highest k' for uridine compared with BEH Amide and BEH HILIC phases.
• Distinct selectivity: Enhanced separation of methylene, hydroxyl, isomeric and ionic pairs.
• Reproducibility: RSD ≤ 2.2 % for relative retention factors across 17 batches.
• pH stability: Stable from pH 2 to 10; < 6 % efficiency loss and modest retention shifts after base exposure.
• Metal-sensitive analytes: MaxPeak HPS hardware provided sharp, symmetric peaks for ATP, ADP and AMP.

Benefits and Practical Applications

• Versatile retention of neutral polar compounds
• Complementary selectivity to other BEH-based HILIC chemistries
• Wide pH range compatibility (2–10) for method development
• Robust reproducibility suitable for QA/QC workflows
• Improved performance for phosphorylated and acidic analytes

Future Trends and Applications

Integration of hybrid zwitterionic phases with ultra-high pressure and low adsorption hardware is expected to advance applications in high-throughput metabolomics, biopharmaceutical quality control and environmental monitoring. Further work on surface modifications may extend compatibility with extreme pH and complex biological matrices.

Conclusion

The Atlantis Premier BEH Z-HILIC phase combines zwitterionic retention, broad pH stability, high efficiency and reproducible performance, making it a valuable tool for a wide range of polar analyte separations. Its compatibility with MaxPeak HPS hardware further enhances analysis of metal-sensitive compounds.

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