Recovery of Metal-Sensitive Analytes on the Arc Premier Solution: System-to-System Reproducibility and a Multi-System Comparison to Conventional LC

Significance of the Topic

Phosphorylated and other metal-sensitive compounds often adsorb to metal surfaces in conventional LC systems, leading to poor recovery, low sensitivity, and irreproducible results. Reliable analysis of these analytes is critical in pharmaceutical, biotechnology, and environmental monitoring where detection of nucleotides, phosphorylated steroids, and related molecules at trace levels is required.

Study Objectives and Overview

This study evaluates the Arc Premier Solution combining ACQUITY Arc with MaxPeak High Performance Surfaces (HPS) technology on multiple instruments to assess system-to-system reproducibility and compare performance against conventional stainless-steel LC. Phosphorylated analytes including ATP, ADP, adenosine, and various phosphorylated steroids were analyzed without system passivation or ion-pair reagents.

Methodology and Instrumentation

• Systems: Three Arc Premier QSM-R and FTN-R instruments with MaxPeak HPS and comparable conventional LC systems.
• Analytes: Adenosine, adenosine diphosphate (ADP), adenosine triphosphate (ATP); hydrocortisone phosphate, dexamethasone sodium phosphate, dexamethasone, dexamethasone acetate.
• Columns: XSelect Premier HSS T3 XP and XBridge Premier BEH C18, 2.5 µm, 4.6×50 mm.
• Detectors: UV/Vis at 260 nm (2489) or PDA (2998).
• Chromatographic conditions: Gradient elution with ammonium acetate or formate buffers; flow rate 1.5 mL/min; column at 40 °C; sample at 10 °C.
• Data analysis: 36–100 injections per system; %RSD calculations and t-tests for statistical comparison.

Main Results and Discussion

• Arc Premier systems showed outstanding reproducibility: %RSD ≤0.28% for metal-sensitive analytes and ≤0.26% for non-metal-sensitive compounds across 36 injections per system.
• Conventional LC systems exhibited broader variability (12–38% RSD) for phosphorylated analytes across instruments.
• Sensitivity at low concentrations was markedly better on Arc Premier: conventional LC often failed to detect low-level ATP/ADP peaks, while Arc Premier maintained linear response from 1–200 µg/mL.
• Statistical analysis confirmed significant improvements in precision for ATP (63-fold) and ADP (41-fold) on Arc Premier (p < 0.05).

Benefits and Practical Applications

• Consistent results across instruments reduce system setup and method qualification time.
• Improved recovery of phosphorylated and charged analytes without passivation or ion-pair reagents.
• Seamless integration into existing HPLC workflows and compatibility with common detection modes facilitates routine QA/QC and research applications.

Future Trends and Opportunities

Advancements in surface coatings and column chemistries will further mitigate metal-analyte interactions. Coupling with high-resolution mass spectrometry and automated platforms may expand applications in proteomics, metabolomics, and biopharmaceutical analysis.

Conclusion

The Arc Premier Solution with MaxPeak HPS technology delivers superior system-to-system reproducibility and enhanced sensitivity for metal-sensitive analytes compared to conventional LC. By minimizing non-specific adsorption, it streamlines workflows and ensures reliable results for challenging compounds.

References

• Li Z.; Shah D.; Dovell A.; Martin W.; Grzonka C.; Dyke J. Challenge Accepted: Arc Premier System Increases Sensitivity and Reproducibility for Hard-to-See Compounds. Waters Application Note 720007267EN, June 2021.