Using a Systematic Screening Protocol and MaxPeak™ HPS Technology to Develop a UHPLC Method for the Analysis of Deferoxamine and its Forced Degradation Products

Significance of the topic

Forced degradation studies are essential for assessing the stability and purity of pharmaceutical agents. Developing a rapid and reliable UHPLC method for deferoxamine and its degradation products supports quality control and regulatory compliance in drug development.

Objectives and study overview

The goal was to apply a systematic screening protocol combined with MaxPeak HPS technology to develop in two days a UHPLC method capable of separating deferoxamine mesylate, its counter ion, and six forced degradation products under acidic and basic hydrolysis conditions.

Methodology and instrumentation

Deferoxamine mesylate was degraded separately with 0.1 N HCl and 0.1 N NaOH at 70 C for one hour and combined. A structured approach began with pH scouting on a BEH C18 column, followed by column and strong solvent screening across four chemistries using low pH formic acid mobile phase. MaxPeak HPS treated flow paths minimized analyte loss to metal surfaces. An ACQUITY Premier quaternary system with QDa mass detector and Empower 3 software managed data acquisition and scheduled SIM detection.

Used Instrumentation

• ACQUITY Premier Quaternary Solvent Manager
• Sample Manager Flow Through Needle and Column Managers
• QDa Mass Detector
• Empower 3 Chromatography Software
• XBridge Premier BEH C18 Column 2.1 x 50 mm 2.5 µm
• XSelect Premier CSH Phenyl Hexyl Column 2.1 x 50 mm 2.5 µm
• XSelect Premier HSS PFP Column 2.1 x 50 mm 2.5 µm
• Atlantis Premier BEH C18 AX Column 2.1 x 50 mm 2.5 µm

Main results and discussion

pH scouting showed superior retention at low pH. Column and solvent screening revealed that the HSS PFP column with a methanol gradient and 0.1% formic acid achieved baseline separation of eight components. USP resolution values exceeded 1.5 and tailing factors ranged from 0.8 to 1.2. A 5 to 95 methanol gradient over 4.9 minutes and scheduled SIRs enhanced sensitivity and shortened run time.

Benefits and practical applications

• Baseline separation of deferoxamine, mesylate counter ion, and six degradants
• Method developed and optimized in under two days
• Structured screening reduces subjective decision making
• MaxPeak HPS treatment prevents analyte adsorption on metal surfaces

Future trends and opportunities

Incorporating software guided design of experiments could further streamline screening. Advances in system and column surface modifications will enhance analysis of metal sensitive compounds. The protocol may be extended to other forced degradation and stability studies in pharmaceutical analysis.

Conclusion

A systematic screening protocol with MaxPeak HPS technology enabled rapid, reproducible development of a UHPLC method for deferoxamine mesylate forced degradation products. Standardized workflows improve documentation and reproducibility across laboratories.

References

• Boissel C Walter TH Shiner SJ ACQUITY Premier Solution Improves UPLC MS Analysis of Deferoxamine
• Maziarz M McCarthy Wrona Systematic Screening Protocol Application Note
• Berthelette KD Nguyen JM Turner Method Development of Antibiotics Using Screening Protocol
• Wyndham KD Walter Iraneta Review of Hybrid BEH Particle Technology