Batch-to-Batch Robustness of MaxPeak Premier Columns for the Analysis Dexamethasone Phosphate and Related Compounds

Importance of the Topic

Analytical methods that yield consistent separation of metal-sensitive compounds are vital for pharmaceutical quality control. Dexamethasone phosphate and related corticosteroids can adsorb to metal surfaces, impairing accuracy. Robust chromatography columns minimize unwanted interactions, ensuring reliable outcomes over long-term operation.

Objectives and Study Overview

This study evaluates batch-to-batch reproducibility of MaxPeak Premier XBridge BEH C18 and XSelect HSS T3 columns. Both variations of the construction (surface treatment) material and the packing media were tested using an established UHPLC method to analyze dexamethasone phosphate, hydrocortisone phosphate, and three related compounds. Emphasis was placed on retention behavior, critical pair resolution, and peak area consistency across column lots.

Methodology and Instrumentation

A UHPLC system (Arc Premier) equipped with a quaternary solvent manager, sample manager, column manager, PDA detector, and QDa mass detector was employed. Key conditions included:

• Columns: MaxPeak HPS XSelect HSS T3 (4.6×100 mm, 2.5 µm) and MaxPeak HPS XBridge BEH C18 (4.6×100 mm, 2.5 µm)
• Mobile phase: 10 mM ammonium formate (A) and acetonitrile with 0.1% formic acid (B)
• Gradient: 10–90% B over 5 or 15 min, 2 min hold, flow 0.5 mL/min
• Temperature: column 35 °C, sample 10 °C
• Injection: 3 µL; detection at 254 nm and MS (ESI+, 100–500 Da)

Main Results and Discussion

Batch-to-batch variability was assessed through three primary metrics:

• Relative retention time: All analytes on both column types showed %RSD ≤ 0.1% across different surface and packing batches.
• Critical pair resolution: Betamethasone phosphate and dexamethasone phosphate were consistently separated with RSD ≤ 0.4% for all column batches.
• Peak area reproducibility: Peak area %RSD ranged from 0.1% to 5.6% for all compounds, demonstrating high quantitative precision.

These consistent results highlight the precision of the MaxPeak High Performance Surfaces manufacturing process and the stability of the packing materials.

Benefits and Practical Applications

• Eliminates revalidation risk by ensuring column uniformity across production lots.
• Enhances method robustness for long-term pharmaceutical analyses.
• Improves analyte recovery and peak shapes for metal-sensitive compounds.

Future Trends and Opportunities

Ongoing development in surface treatments and hybrid particle technologies will further suppress unwanted interactions, enabling lower detection limits and wider applicability in bioanalysis and environmental testing. Integration with advanced data analytics and high-throughput workflows could drive new applications in quality-by-design method development.

Conclusion

MaxPeak Premier XSelect HSS T3 and XBridge BEH C18 columns exhibit exceptional batch-to-batch reproducibility for critical chromatographic parameters. Their robust performance supports reliable analysis of dexamethasone phosphate and related compounds, simplifying long-term method maintenance and regulatory compliance.

Reference

• Lauber THW et al. Low Adsorption UPLC Columns Based on MaxPeak High Performance Surfaces. Waters White Paper. 2020.
• Alkhateeb FL, Rainville PD. Analytical Quality by Design Based Method Development for Dexamethasone Phosphate and Related Compounds Using Arc Premier HPS Technology. Waters Application Note. 2021.
• Collins KE et al. Stainless Steel Surfaces in LC Systems, Part II. LC GC North America. 2000.