Analysis of Urea

Significance of the Topic

Urea is a small, polar molecule of great importance in clinical diagnostics, pharmaceutical manufacturing, environmental monitoring and food quality control. Accurate and rapid quantification of urea supports process control in industrial applications, clinical assessments of renal function and ensures product safety in various sectors.

Objectives and Overview

This application note demonstrates a hydrophilic interaction liquid chromatography (HILIC) method using the Shim-pack GIST Amide column to achieve robust separation and quantification of urea. The study highlights method performance, simplicity of operation and suitability for routine quality control.

Methodology and Used Instrumentation

The analysis was carried out under isocratic conditions on a Nexera XR UHPLC system with the following components and settings:

• Column: Shim-pack GIST-HP Amide, 150 mm × 2.1 mm I.D., 1.9 µm particle size
• Column oven temperature: 40 °C
• Mobile phase: Water (A) / Acetonitrile (B) 15/85 (v/v), isocratic mode
• Flow rate: 0.25 mL/min
• Injection volume: 1 µL
• Detection: UV at 200 nm
• Sample: Urea standard (5 mg/mL) in water/acetonitrile (15/85 v/v)

Main Results and Discussion

The HILIC method yielded a single, well defined urea peak eluting at approximately 2.2 minutes with good symmetry and minimal tailing. The baseline was stable throughout the 6-minute run time. The rapid analysis, combined with high retention and clear resolution, confirms method suitability for precise quantification without derivatization.

Benefits and Practical Applications

The described HILIC approach offers:

• Fast turnaround: total run time under 6 minutes
• High sensitivity: reliable detection at low concentrations
• Simplified sample preparation: direct injection of standard solutions
• Compatibility with routine QC workflows in pharmaceutical, clinical and environmental laboratories

Future Trends and Possibilities

Further enhancements may include coupling the HILIC method with mass spectrometric detection to improve sensitivity and selectivity, extension to urea-related metabolites, integration into automated high-throughput platforms and exploration of greener solvent systems to reduce environmental impact.

Conclusion

The Shim-pack GIST Amide HILIC method on a Nexera XR system provides a rapid, robust and sensitive solution for urea analysis, suitable for diverse applications in research and quality control laboratories.

References

• Shimadzu Corporation. 2022. Application Note ERAS-1000-0007A. LC Hydrophilic Interaction (HILIC) Shim-pack Series: Analysis of Urea. First Edition, Sep. 2022.