HPLC Method for Separation of Thiourea, Thiouracil (TU), Methylthiouracil (MTU) and Propylthiouracil (PTU) on Primesep P Column

Importance of the Topic

Thiourea and its derivatives are sulfur-containing compounds widely used in pharmaceutical and industrial contexts. Effective separation and analysis of these molecules are essential for quality control in drug development and process monitoring.

Objectives and Study Overview

This study aimed to develop a rapid and reliable isocratic HPLC method for separating thiourea, thiouracil, methylthiouracil, and propylthiouracil using a mixed-mode stationary phase.

Methodology and Instrumentation

The method employed a Primesep P column under isocratic conditions with UV detection. Key parameters included:

• Column: Primesep P, 4.6 × 250 mm, 5 µm
• Mobile phase: acetonitrile/water (5/95) with 0.1% phosphoric acid
• Flow rate: 1.0 mL/min
• Detection: UV at 200 nm

Main Results and Discussion

The optimized method achieved baseline resolution of all four analytes within approximately 12 minutes. The mixed-mode cation-exchange and π–π interactions of the Primesep P phase provided sharp peaks and consistent retention, demonstrating robustness and reproducibility.

Benefits and Practical Applications

This simple isocratic approach simplifies mobile phase preparation and allows straightforward implementation in pharmaceutical quality control and research laboratories. The method can be extended to other polar or ionizable compounds.

Future Trends and Applications

Enhancements may include shorter columns and smaller particle sizes for faster separations, adoption of core-shell particles for improved efficiency, and coupling with mass spectrometry for increased selectivity and sensitivity.

Conclusion

An efficient and reliable HPLC method was established for thiourea derivatives using a mixed-mode Primesep P column. The approach offers rapid analysis, ease of use, and adaptability for various polar compounds.

References

1. SIELC Technologies. HPLC Method for Separation of Thiourea, Thiouracil, Methylthiouracil and Propylthiouracil on Primesep P Column; Application Note CoAp #2023-3; 2023.