Supercritical fluid chromatography is not (only) normal-phase chromatography

Supercritical fluid chromatography (SFC) is a mature chromatographic technique, as the first experiments were presented over 60 years ago [1], and a large body of literature exists describing fundamentals [2,3] and ever-increasing applications [4]. Its potential for enantioresolution [5,6] and its interest for preparative-scale separations [7,8] were acknowledged a long time ago [9], but its capabilities for achiral and analytical-scale separations were re-discovered and more widely recognized only in the past decade [10]. This is due to the introduction of high-performance chromatographic instruments by several manufacturers, finally meeting the expectations of chromatographers in terms of repeatability [11], robustness, sensitivity, capability to transfer a method from one laboratory and instrument to another one [12,13] and hyphenation to mass spectrometry [14], [15], [16]. Although significant efforts have been made to educate chromatographers to a better use of the technique [17], some misconceptions are often expressed in published papers. The most frequent one is the supposed “normal-phase” (NP) operating mode. Indeed, very frequently, SFC is simply considered as a normal-phase technique, based on different arguments. The possible reasons expressed for this NP behaviour are based on the non-polar nature of the major mobile phase component (carbon dioxide), the observed elution order of analytes depending on their polarity, the nature of the stationary phases most employed, or finally, the supposed adsorption retention mechanism. However, considering SFC only through this angle could be deceiving, as one would expect similar chromatograms when transferring a method from NP-liquid chromatography to SFC. In addition, it would be greatly reducing the application scope of the technique. This is what we will demonstrate in this paper, considering all usual arguments to support the supposed normal-phase mode.

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