Improving Metal-Sensitive Analyte Recovery on Various LC Systems Using MaxPeak Premier Columns

Importance of Topic

Metal-induced adsorption in liquid chromatography (LC) compromises the accuracy and sensitivity of analyses for acidic probes such as phosphorylated peptides and nucleotide analogs. Addressing these interactions is essential for reliable quantitation in proteomics, metabolomics, and pharmaceutical quality control.

Study Aims and Overview

This application note investigates whether MaxPeak Premier columns, featuring high-performance surface (HPS) technology, can improve recovery of the metal-sensitive analyte adenosine 5’-(α,β-methylene) diphosphate (AMPcP) on three non-Waters LC platforms. By comparing peak areas of AMPcP against an insensitive comparator (adenosine), the study aims to demonstrate system-agnostic benefits of surface-passivated column hardware.

Methodology and Instrumentation

The AMPcP/adenosine standard was prepared in Milli-Q water and analyzed under transferred gradient conditions across three LC systems. Key instrumentation included:

• Agilent 1290 Infinity I with DAD detection at 260 nm
• Shimadzu Nexera-I LC2040 with UV detection at 260 nm
• Thermo Vanquish UHPLC with UV detection at 260 nm

Columns evaluated:

• XSelect HSS T3 XP Column, 2.1 × 50 mm, 2.5 µm
• XSelect Premier HSS T3 Column, 2.1 × 50 mm, 2.5 µm (MaxPeak Premier HPS)
• Zorbax SB-Aq Column, 2.1 × 50 mm, 3.5 µm

All analyses were performed at 40 °C with a 0.36 mL/min flow (0.70 mL/min on Shimadzu), using ammonium acetate buffers (pH 6.8) in water/acetonitrile. Data acquisition employed Empower 3 or Chromeleon 7.2 software.

Main Results and Discussion

Comparative peak area and peak area ratio (AMPcP/adenosine) results across systems:

• Agilent 1290: Premier column yielded >50 % higher AMPcP peak area and raised the area ratio from ~0.35 to 0.54 versus steel columns.
• Shimadzu Nexera-I: Despite increased injection volume, Premier column improved AMPcP area by ~10 % and raised the ratio by ~0.2 units.
• Thermo Vanquish: Premier column showed ~20 % higher AMPcP peak area and similar gain in the area ratio.

Across all platforms, adenosine signals remained constant, confirming that improvements derive from reduced metal adsorption rather than changes in chromatographic selectivity or sample load.

Benefits and Practical Applications

• Enhanced analyte recovery for acidic, metal-sensitive compounds without surface passivation reagents.
• Improved peak shape and sensitivity translate to lower detection limits and more accurate quantitation.
• Compatibility with diverse LC configurations simplifies method transfer and instrument utilization in pharmaceutical and bioanalytical laboratories.

Future Trends and Opportunities

Further developments may include extension of HPS technology to a broader range of stationary phases and system components, coupling with mass spectrometry for ultra-trace analysis, and exploration of long-term robustness for challenging sample matrices. Adoption of inert hardware across the LC ecosystem promises enhanced data quality in omics workflows and regulated environments.

Conclusion

MaxPeak Premier columns deliver system-agnostic improvements in recovery and quantitation of metal-sensitive analytes, as demonstrated by significant gains in AMPcP peak area across three non-Waters LC systems. This universally applicable solution supports more reliable analyses without additional surface treatments.

References

1. Jung M., Lauber M. Demonstrating Improved Sensitivity and Dynamic Range with MaxPeak High Performance Surface (HPS) Technology: A Case Study in the Detection of Nucleotides. Waters Application Note; 2020.
2. Patel A., Simeone J., Delano M., Dyke J., Rzewuski S., Jung M., Shiner S. Premier Standards to Investigate the Inertness of Chromatographic Surfaces. Waters Application Note; 2021.
3. DeLano M., Walter T.H., Lauber M., Gilar M., Jung M.C., Nguyen J., Boissel C., Patel A.V., Bates-Harrison A., Wyndham K. Using Hybrid Organic–Inorganic Surface Technology to Mitigate Analyte Interactions with Metal Surfaces in UHPLC. Analytical Chemistry. 2021;93(22):5773–5781.
4. Isaacs G., Plumb R. ACQUITY Premier LC Technology Significantly Improves Sensitivity, Peak Shape, and Recovery for Phosphorylated and Carboxylate Lipids. Waters Application Note; 2021.
5. Smith K., Rainville P. Improved Analytical Sensitivity and Chromatographic Peak Shape for the Quantitation of TCA Cycle Analytes in Human Plasma using the ACQUITY Premier System Solution. Waters Application Note; 2021.
6. Waters Corporation. AMPcP and Adenosine Standard. Care and Use Manual; 2021.
7. Lauber M., Walter T.H., Gilar M., Delano M., Boissel C., Smith K., Rainville P., Belanger J., Wyndham K. Low Adsorption HPLC Columns Based on MaxPeak High Performance Surfaces. Waters White Paper; 2020.