Analysis of thiopurines using aqueous normal phase chromatography

Area of Science:

• Analytical Chemistry
• Chromatography

Background:

• Thiopurines are important pharmaceutical compounds.
• Chromatographic methods are essential for their analysis.
• Aqueous normal phase (ANP) chromatography offers unique separation capabilities.

Purpose of the Study:

• To investigate the chromatographic behavior of thiopurines.
• To evaluate the effectiveness of aqueous normal phase (ANP) conditions.
• To assess the suitability of silica hydride-based columns for thiopurine separation.

Main Methods:

• Utilized aqueous normal phase (ANP) chromatography.
• Employed a silica hydride-based stationary phase.
• Tested both isocratic and gradient elution modes.
• Employed UV detection for high concentration samples and mass spectrometry (MS) for low concentration samples.

Main Results:

• Demonstrated the feasibility of ANP chromatography for thiopurine analysis.
• Evaluated the performance of isocratic and gradient elution methods.
• Assessed the repeatability of successive runs, particularly for gradient methods.
• Provided insights into the stationary phase equilibration time in gradient elution.

Conclusions:

• Aqueous normal phase (ANP) chromatography on silica hydride columns is a viable method for thiopurine analysis.
• Both UV and MS detection are suitable depending on sample concentration.
• Gradient elution methods require careful consideration of stationary phase equilibration for reproducible results.