Development of a Generic Gradient SFC Method for Application in Drug Discovery and Development

In recent years, packed column Supercritical Fluid Chromatography (SFC) has received renewed interest within the pharmaceutical industry. Its inherent features such as speed, efficiency and versatility make SFC the perfect complement to reversed-phase liquid chromatography (RP-LC) and hydrophilic interaction liquid chromatography (HILIC) in drug discovery and development. Moreover, the ‘green’ credentials of the technique are becoming more and more appreciated. Research in SFC is principally directed towards the development of novel stationary phases and generic method development workflows.

However, the pharmaceutical industry is looking at SFC with growing interest but specifically requires instrumentation that meets ever-increasing regulatory requirements. This article presents a generic gradient method applicable to the separation of analytes with a wide polarity range, the separation of complex mixtures and the trace level determination of impurities utilising the common 2-ethylpyridine and cyanopropyl SFC stationary phases.

The mobile phase composition is such that several detection types including mass spectrometry (MS) may be applied. The performance of current state-of-the-art SFC instrumentation is illustrated with a mixture composed of several compounds of pharmaceutical interest and through the determination of thiourea in a pharmaceutical intermediate at the 0.01% (w/w) level.