Transfer the USP Method for Ceftizoxime from a Traditional 5-μm Column to an Agilent Poroshell 120

Transfer of USP Method for Ceftizoxime to Agilent Poroshell 120 Columns

Significance of the Topic

The transfer of pharmacopeial assays to advanced column technologies can drastically reduce analysis time and solvent consumption while maintaining compliance with regulatory standards.

Study Objectives and Overview

This application note aimed to evaluate the feasibility of migrating the USP monograph method for ceftizoxime from a conventional 5 µm C18 column to Agilent Poroshell 120 superficially porous columns with 2.7 µm particle size. Performance metrics were compared against USP Chapter 621 requirements.

Methodology and Instrumentation

• Chromatographic system: Agilent 1200 Infinity Series Rapid Resolution LC with binary pump, autosampler, column compartment, and diode array detector.
• Columns tested:
  • ZORBAX Eclipse Plus C18, 4.6×250 mm, 5 µm
  • Poroshell 120 EC-C18, 4.6×100 mm and 4.6×75 mm, 2.7 µm
• Mobile phase: 90:10 buffer (pH 3.6, citric acid/sodium phosphate) to acetonitrile.
• Flow rate: 2.0 mL/min; Injection volumes: 10 µL, 4 µL, 3 µL.
• Detection: UV at 254 nm; Column temperature: 25 °C.

Results and Discussion

The traditional 5 µm column achieved a resolution of 22.6 in 12 minutes, well above the USP minimum of 4.0. Switching to a 4.6×100 mm Poroshell 120 column reduced analysis time to 4 minutes and improved resolution to 25.2. A shorter 4.6×75 mm variant maintained compliance under USP guidelines. Column-to-column reproducibility studies showed consistent retention times and resolution across different batches.

Benefits and Practical Application

• Time savings: Reduction of run times from 12 minutes to as low as 4 minutes.
• Solvent consumption: Over 65 percent decrease in mobile phase usage per analysis.
• Compatibility: Lower backpressure allows use on standard HPLC systems.
• Regulatory compliance: All USP system suitability requirements met for ceftizoxime assay.

Future Trends and Potential Applications

Advancements in superficially porous particle technology are expected to drive wider adoption in pharmaceutical quality control, including high-throughput environments. Integration with ultra-high-performance systems and digitized workflows can further enhance efficiency and data robustness.

Conclusion

The study confirms that the USP assay for ceftizoxime can be effectively migrated from a 5 µm column to Agilent Poroshell 120 superficially porous columns. This transition offers substantial time and cost efficiencies without compromising chromatographic performance or pharmacopeial compliance.

References

1. A. Mack. USP Analysis of Diphenhydramine and Pseudoephedrine Using an Agilent Poroshell 120 EC-CN Column. Agilent Technologies 2013, publication number 5991-1687EN.
2. United States Pharmacopeia. Ceftizoxime. USP35-NF30 Chapter 2574. Rockville MD 2012.