Consumables, LC/MS, LC columns
IndustriesManufacturerWaters
Importance of the topic
High sensitivity and reproducible recovery of analytes are essential for reliable LC-MS analysis, especially at trace concentrations. Negative interactions between analytes and column hardware can lead to sample loss and variable results. The development of inert high performance surfaces addresses these challenges by reducing analyte binding to metal surfaces and improving overall method performance.
Goals and overview of the study
This work evaluates the impact of integrating MaxPeak High Performance Surfaces (HPS) into the mixed-mode Atlantis PREMIER BEH C18 AX column. The aim is to quantify improvements in analyte recovery, sensitivity, and reproducibility for diverse compound classes under reversed-phase and anion exchange conditions.
Methodology and instrumentation
A series of standard solutions containing phosphorylated metabolites, pharmaceutical agents, and biological markers were separated using the Atlantis PREMIER BEH C18 AX stationary phase. Comparative analyses were performed with and without MaxPeak HPS column hardware. Gradient elution employed aqueous ammonium acetate or formate buffers at pH 3.0 or 9.2 in acetonitrile mixtures over short run times. Peak areas were recorded by LC-MS.
Used Instrumentation
- Waters ACQUITY UPLC system coupled to a mass spectrometer
- Atlantis PREMIER BEH C18 AX column
- MaxPeak High Performance Surfaces column hardware
Main results and discussion
Integration of MaxPeak HPS hardware yielded significant enhancements in signal intensity: AMP peak area increased by 1.5-fold and ATP by 19-fold under alkaline gradient conditions. Four additional compounds spanning different chemical functionalities exhibited area gains of 1.3- to 1.6-fold. Enhanced recovery was attributed to minimization of metal binding interactions for phosphorylated species and sulfur- or nitrogen-containing drugs.
Benefits and practical applications of the method
- Improved sensitivity for low-abundance analytes, facilitating trace analysis
- Greater reproducibility across runs by limiting surface adsorption
- Simplified method development for polar and ionizable compounds
Future trends and potential applications
Extension of inert surface technology to other column chemistries may further enhance LC-MS performance for diverse analytes. Emerging bioanalytical demands for metabolomics and pharmacokinetics will benefit from robust hardware that reduces sample loss and extends column lifetime.
Conclusion
Incorporation of MaxPeak HPS into the Atlantis PREMIER BEH C18 AX column substantially increases analyte response and method robustness. This advancement supports sensitive quantitation across a range of compounds and offers a practical solution to mitigate analyte-surface interactions in LC-MS workflows.
References
1. Sakamaki H et al Evaluation of column hardware on liquid chromatography-mass spectrometry of phosphorylated compounds J Chrom A 2015 1381 125-131
2. Walter TH et al A New Mixed-Mode Reversed Phase Anion Exchange Stationary Phase Based on Hybrid Particles Waters Corporation Application Note 2020
3. Tuytten R et al Stainless-steel electrospray probe A dead end for phosphorylated organic compounds J Chrom A 2006 1104 209-221
4. Schenk WA The coordination chemistry of small sulfur-containing molecules a personal perspective Dalton Trans 2011 40 1209-1219
5. Hancock RD Martell AE Ligand design for selective complexation of metal ions in aqueous solution Chem Rev 1989 89 1875-1914
6. Chen AY et al Targeting metalloenzymes for therapeutic intervention Chem Rev 2019 119 1323-1455
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