Rapid Analysis of Cephalosporins and Related Drug Substances Using CORTECS™ Premier Columns featuring MaxPeak™ Technology

Significance of the Topic

Cephalosporins are essential antibiotics used to treat serious bacterial infections, including strains resistant to other agents. Rapid, precise, and accurate analytical methods are critical in pharmaceutical quality control to ensure drug safety, potency, and batch consistency.

Objectives and Study Overview

This work develops and evaluates a rapid reversed-phase liquid chromatography (RPLC) method for common cephalosporins, leveraging CORTECS Premier Columns with MaxPeak High Performance Surfaces (HPS) Technology. The method is benchmarked against a traditional stainless-steel chromatography setup in terms of speed, sensitivity, reproducibility, and quantitative performance.

Methodology and Instrumentation

Sample Preparation:

• Individual stock solutions of cefapirin, cefaclor, and cephalexin at 1 mg/mL in water
• Mixed standard at 20 µg/mL for precision testing
• Cephalexin calibration standards prepared at 1–100 µg/mL for linearity assessment

Instrument Setup:

• ACQUITY Premier System with CORTECS Premier Columns featuring MaxPeak HPS Technology
• ACQUITY UPLC I-Class PLUS System with stainless-steel hardware for comparison
• 2998 Photodiode Array Detector for UV detection
• Empower 3 Software for data acquisition and processing

Chromatographic Conditions:
The method employs a rapid gradient elution under acidic mobile phase conditions, achieving complete separation in under two minutes per injection.

Key Results and Discussion

• Precision: Ten replicate injections of the cephalosporin mix yielded retention time and peak area RSDs below 5%, demonstrating high repeatability.
• Linearity: The cephalexin calibration curve (1–100 µg/mL) produced R² > 0.999, confirming suitability for quantitative analysis.
• System Comparison: The Premier setup showed up to 40% greater peak height, improved peak symmetry, and enhanced retention compared to stainless-steel hardware, indicating reduced metal–analyte interactions.
• Accuracy: A cephalexin drug sample at 50 µg/mL was quantified at 49.27 µg/mL (98% recovery), supporting method accuracy for routine QC.

Benefits and Practical Applications

• Sub-two-minute run times significantly increase sample throughput in quality control laboratories.
• MaxPeak HPS Technology mitigates undesirable metal interactions, enhancing sensitivity and chromatographic performance for acidic analytes.
• Proven precision, linearity, and accuracy support reliable routine analysis and batch release testing of cephalosporin drug substances.

Future Trends and Potential Applications

• Extension of high-performance surface technologies to other acidic pharmaceuticals and complex biologics.
• Integration with mass spectrometry detection for enhanced selectivity and trace-level quantitation.
• Development of multiplexed, high-throughput workflows for impurity profiling and stability studies.
• Advances in method miniaturization and green solvent protocols to reduce sample and solvent consumption.

Conclusion

The use of CORTECS Premier Columns with MaxPeak HPS Technology offers a robust, rapid RPLC solution for cephalosporin analysis, delivering high precision, excellent linearity, and reliable accuracy in under two minutes, outperforming traditional stainless-steel chromatography setups and meeting stringent quality control requirements.

Reference

• Bui T, Preuss CV. Cephalosporins. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2020.
• Walter T, Trudeau M, Simeone J, et al. Low Adsorption UPLC Systems and Columns Based on MaxPeak High Performance Surfaces: The ACQUITY Premier Solution. Waters Corporation; May 2021. Application Note 720007128.
• Layton C, Rainville P. Improvements in Chromatographic Performance for Stability Indicating Methods of Antiviral Drugs With MaxPeak Premier Technology. Waters Corporation; Jan 2022. Application Note 720007487.
• Shah D, Smith K, Yang J, Hancock P. Analysis of Fourteen Organic Acids in Various Beverages Using the ACQUITY UPLC H-Class PLUS and ACQUITY QDa Mass Detector. Waters Corporation; Aug 2021. Application Note 720007289.
• Waters Corporation. CORTECS Columns, Applications Notebook. Waters Corporation; 2012. Document 720004739.