Hydrophilic Interaction Liquid Chromatography (HILIC) Method Migration Part 1: From Legacy HPLC Systems to the Alliance™ iS HPLC System

Importance of the topic

Hydrophilic interaction liquid chromatography HILIC is widely adopted in regulated laboratories for analyzing polar compounds using high organic mobile phases. Ensuring that established HILIC methods can be transferred intact to modern HPLC platforms is essential to maintain data consistency, reduce revalidation effort, and meet regulatory requirements.

Objectives and study overview

This work evaluates migration of the United States Pharmacopeia USP monograph for cetirizine hydrochloride assay and its organic impurities from two legacy HPLC systems to the Alliance iS HPLC System. Key system suitability parameters—tailing factor, resolution, signal to noise ratio, peak area repeatability, and retention time repeatability—were assessed to verify comparable performance across platforms.

Methodology and instrumentation

The USP methods employ isocratic separations of 10 minutes for the assay and 18 minutes for impurity profiling on an XBridge BEH HILIC column at 25 °C. The mobile phase consisted of acetonitrile water and sulfuric acid in a 93 6.6 0.4 ratio. Standards for the assay and impurity solutions were prepared at relevant concentrations. Three HPLC configurations were used: two legacy systems with UV detectors at 230 nm and the Alliance iS system with dual wavelength detection. Empower 3 software managed data processing and custom calculations.

Instrumentation used

• Legacy System 1 with 2489 UV Vis detector at 230 nm, 10 points per second
• Legacy System 2 with variable wavelength detector at 230 nm, 6.87 Hz
• Alliance iS HPLC System with dual wavelength UV detector at 230 nm, 10 points per second
• XBridge BEH HILIC column, 130 Å pore size, 5 µm particle size, 4.6 × 250 mm
• Empower 3 chromatography data system

Major results and discussion

All systems met USP criteria for the cetirizine assay, achieving peak area RSD below 0.73 and tailing factor below 2.0. The Alliance iS delivered the lowest area RSD of 0.042 . In impurity analysis, resolution and tailing factor were comparable, while the Alliance iS exhibited superior signal to noise due to enhanced needle wash routines that eliminated carryover seen on one legacy instrument. Custom intrasample calculations further confirmed accurate signal to noise assessment when blanks were free of interfering peaks.

Benefits and practical applications

• Minimized revalidation workload by transferring existing HILIC methods
• Improved injection precision and sensitivity on the Alliance iS system
• Reduced carryover through advanced wash design
• Consistent chromatographic performance across platforms

Future trends and potential applications

Future HPLC developments are expected to focus on further enhancements in wash protocols and sensitivity for trace analysis. Automated method transfer workflows and predictive diagnostics may streamline migration of complex HILIC assays. Extending these capabilities to mass spectrometry interfaces could broaden applications in pharmaceutical impurity profiling.

Conclusion

The Alliance iS HPLC System successfully adopted the USP cetirizine hydrochloride HILIC methods from legacy platforms, meeting or surpassing performance in precision and sensitivity. Its optimized needle wash mechanism underpins improved data quality and supports reliable method migration in regulated environments.

References

1. United States Pharmacopeia USP monograph for Cetirizine Hydrochloride assay and organic impurities Rockville MD USP 01 Sep 2021
2. United States Pharmacopeia General Chapter 621 Chromatography Rockville MD USP 01 Apr 2023
3. Waters support article How to Create an Intersample Custom Field Empower Tip 93 Waters 11 Nov 2022