High Speed Analysis of Pregabalin in Accordance with Chapter 621 in USP 40

Importance of the Topic

The United States Pharmacopeia (USP) monographs have traditionally relied on 5 µm HPLC columns, leading to lengthy run times, high solvent consumption and increased operating costs. Adapting these methods with smaller particle sizes and optimized flow rates in compliance with USP General Chapter <621> offers a route to faster, more economical analyses while preserving method performance and regulatory compliance.

Objectives and Study Overview

This work aims to demonstrate a high-speed, isocratic HPLC assay for Pregabalin, an antiepileptic agent, based on the USP 40 monograph. By applying the permissible adjustments defined in Chapter 621, three column configurations were evaluated on Shimadzu Prominence-i LC-2030C Plus and Nexera X2 systems to compare retention times and system suitability against the original method.

Methodology

An isocratic mobile phase of water/acetonitrile (95:5, v/v) at 25 °C was used for all experiments. The original method employed a 4.6 × 250 mm, 5 µm column at 1.0 mL/min flow. Two fast methods used columns of 3.0 × 150 mm, 3 µm at 0.7 mL/min and 2.1 × 100 mm, 2 µm at 0.5 mL/min. Column length, internal diameter and flow rates were adjusted following the L/dp ratio constraints (−25 % to +50%) and flow rate scaling equations in USP <621>.

Used Instrumentation

• Shim-pack™ GIST C18 columns: 4.6 × 250 mm, 5 µm; 3.0 × 150 mm, 3 µm; 2.1 × 100 mm, 2 µm
• Shimadzu Prominence-i LC-2030C Plus system with standard UV flow cell (205 nm detection)
• Shimadzu Nexera X2 system with SPD-M30A UV detector (205 nm)

Main Results and Discussion

Fast methods significantly reduced retention times from 9.8 min (original) to 3.7 min (3 µm column) and 1.9 min (2 µm column), with peak tailing factors below 1.5 and RSD values well under USP limits. Chromatographic resolution and system suitability met all acceptance criteria, demonstrating equivalent performance across both Shimadzu platforms.

Benefits and Practical Applications

This approach delivers marked decreases in analysis time and solvent use, improving laboratory throughput and cost efficiency. The method’s compliance with USP <621> ensures easy adoption in quality control environments without method revalidation burdens.

Future Trends and Opportunities

Continued advances in ultra-high-performance columns, core-shell technologies and automated systems will further accelerate pharmaceutical workflows. Integration with mass spectrometry and green chemistry practices promises enhanced selectivity and reduced environmental impact.

Conclusion

The study confirms that applying USP <621> guidelines with smaller-particle, shorter columns on Shimadzu systems achieves rapid, reliable Pregabalin analysis, reducing run times and solvent consumption while maintaining regulatory compliance.

References

• Application News No. L464, Shimadzu
• USP General Chapter 621, USP 40–NF 35
• USP Monograph: Pregabalin, USP 40–NF 35