UHPLC-UV Method for the Determination of Esomeprazole Using a Syncronis C18 1.7 μm Column

Significance of the Topic

Esomeprazole is a widely prescribed proton pump inhibitor used to treat gastroesophageal reflux disease and stomach ulcers. Rapid and reproducible quantification of this active pharmaceutical ingredient is essential in quality control laboratories to ensure product consistency and patient safety. The adoption of ultra-high-performance liquid chromatography with UV detection (UHPLC-UV) facilitates faster analysis times, reduced solvent usage, and improved laboratory throughput.

Objectives and Study Overview

This application note aims to demonstrate a UHPLC-UV method for determining esomeprazole using a Thermo Scientific Syncronis C18 1.7 µm column. Key objectives include achieving a total run time of less than two minutes, excellent peak shape, and high reproducibility across replicate injections.

Methodology

Sample Preparation:

• Working standard: 50 µg/mL esomeprazole in HPLC-grade water.

Chromatographic Conditions:

• Column: Syncronis C18, 1.7 µm, 50 × 2.1 mm.
• Mobile phase A: water with 0.1 % ammonia solution.
• Mobile phase B: acetonitrile.
• Gradient: start at 20 % B, hold 0.5 min; ramp to 70 % B at 2 min; return to 20 % B at 2.01 min; re‐equilibrate until 3 min.
• Flow rate: 0.4 mL/min.
• Column temperature: 30 °C.
• Injection volume: 1 µL (partial loop).
• Injection wash: 80:20 water:acetonitrile.
• Detection wavelength: 302 nm UV.
• System backpressure: ~450 bar.

Instrumentation Used

The analysis was performed on a Thermo Scientific Accela UHPLC system equipped with a UV detector set to 302 nm. The Syncronis C18 column provides high surface area silica, dense bonding, and double endcapping for exceptional reproducibility.

Main Results and Discussion

The method achieved rapid separation of esomeprazole in under two minutes. Key performance metrics from six replicate injections are:

• Retention time: 1.66 min (RSD 0.6 %).
• Peak asymmetry factor: 1.24 (RSD 1.6 %).

Chromatographic profiles demonstrated narrow, symmetric peaks with minimal baseline noise, confirming the column’s suitability for high-throughput assays.

Benefits and Practical Applications

By employing a sub-2 µm UHPLC column and optimized gradient, laboratories can benefit from:

• Reduced analysis time and increased sample throughput.
• Lower solvent consumption and operational costs.
• High reproducibility suitable for routine quality control of esomeprazole formulations.

Future Trends and Opportunities

Future developments may include:

• Coupling UHPLC-UV methods with mass spectrometry for enhanced selectivity and sensitivity.
• Expanding the method to simultaneous determination of related proton pump inhibitors or degradation products.
• Implementing greener mobile phase additives to further reduce environmental impact.

Conclusion

The UHPLC-UV method using the Syncronis C18 1.7 µm column offers a rapid, reliable, and reproducible approach for quantifying esomeprazole in pharmaceutical samples. Its high throughput and robust performance make it an excellent choice for routine quality control operations.