Analytical and semipreparative separation of 25 (R/S)‐spirostanol saponin diastereomers using supercritical fluid chromatography

Abstract

Spirostanol saponins are a class of steroidal saponins with many pharmacological activities. The structural complexity of the spirostanol saponins presents a daunting challenge in separating their 25 R/S diastereomers. Using two CHIRALPAK IC columns coupled in series, six 25 (R/S)-spirostanol saponin diastereomers from the Trigonella foenum-graecum L. seed were successfully separated using supercritical fluid chromatography (SFC) for the first time. In addition, three 25 (R/S)-spirostanol saponin diastereomers were isolated into their respective individual isomers. The structures of the isolated isomers were unambiguously confirmed by NMR analysis. The SFC method development strategy and its associated underlying principles presented in this paper are generally applicable. SFC is a viable addition to the natural product research toolbox, especially for stereoselective analysis and purification.

Simple, rapid, and environmentally friendly method for the separation of isoflavones using ultra‐high performance supercritical fluid chromatography

“…SFC is a separation technique widely used in the field of medicine and food industry for separation of both chiral and achiral compounds . Supercritical state is maintained when carbon dioxide occurs above its critical temperature and pressure.…”

Towards eco‐friendly secondary plant metabolite quantitation: Ultra high performance supercritical fluid chromatography applied to common vervain (Verbena officinalis L.)

“…Consequently, UHPSFC has increasingly developed to a valuable alternative to organic solvent based reversed phase chromatography in modern natural product analysis [13][14][15]. The main potential of UHPSFC is seen as analytical alternative for the assessment of nonpolar and moderate polar plant constituent; its applicability to more polar compounds is still less researched [13][14][15][16][17][18][19][20][21][22]. A second limitation in current literature is the focus of most validated, quantitative applications on single secondary metabolite classes.…”

Large‐scale chiral supercritical fluid chromatography of a key intermediate in the synthesis of two S1P1 final active pharmaceutical ingredients

“…In supporting early phase chemistry programs, SFC using chiral stationary phases (CSP) has proven to be a direct and efficient way to isolate individual stereoisomers and provided small amounts of each enantiomer in high ee for potency tests . In the drug discovery environment, a unique challenge facing separation chemists on a daily basis is not only to resolve a significant number of structurally diverse compounds but also to provide a large quantity of pure stereoisomers for various toxicology studies, which usually requires from tens of grams to several hundred grams.…”

“…small amounts of each enantiomer in high ee for potency tests [1][2][3][4][5][6][7]. In the drug discovery environment, a unique challenge facing separation chemists on a daily basis is not only to resolve a significant number of structurally diverse compounds but also to provide a large quantity of pure stereoisomers for various toxicology studies, which usually requires from tens of grams to several hundred grams.…”