Analysis of thiopurines using aqueous normal phase chromatography

• Thiopurines are highly polar.
• Aqueous normal phase method is described.
• Analysis done without derivatization or ion-pairing.
• Method can be easily adapted to clinical samples.

The chromatography of several thiopurines is investigated using aqueous normal phase (ANP) conditions in conjunction with a silica hydride-based column. Both isocratic and gradient elution modes are tested. Detection of higher concentration samples is done by UV to demonstrate feasibility in this format while lower concentration samples utilize mass spectrometry (MS). Repeatability of successive runs is also tested with particular attention to gradient methods where the equilibration time of the stationary phase can be evaluated.

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