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

Abstract

Isoflavones are natural substances that exhibit hormone-like pharmacological activities. The separation of isoflavones remains an analytical challenge because of their similar structures. We show that ultra-high performance supercritical fluid chromatography can be an appropriate tool to achieve the fast separation of 12 common dietary isoflavones. Among the five tested columns the Torus DEA column was found to be the most effective column for the separation of these isoflavones. The impact of individual parameters on the retention time and separation factor was evaluated. These parameters were optimized to develop a simple, rapid, and green method for the separation of the 12 target analytes. It only took 12.91 min using gradient elution with methanol as an organic modifier and formic acid as an additive. These isoflavones were determined with limit of quantitation ranging from 0.10 to 0.50 μg/mL, which was sufficient for reliable determination of various matrixes.

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

“…In SFC, a decrease of the temperature leads typically to an increase of the mobile phase density and thus to faster elution of the compounds. However, in subcritical conditions with high modifier concentrations, prediction of temperature influence is not that simple and, as reported by different studies, the influence of temperature on the retention time can change during the gradient, related to fluid compressibility changes [20,[35][36][37]. As shown in Figure 2, this trend was observed also in the presented study.…”

“…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.…”

Efficient preparative separation of 6‐(4‐aminophenyl)‐5‐methyl‐4, 5‐dihydro‐3(2H)‐pyridazinone enantiomers on polysaccharide‐based stationary phases in polar organic solvent chromatography and supercritical fluid chromatography

“…SFC is now well known for its fast analysis and green mobile phase that consists of carbon dioxide (CO 2 ) and organic modifier (normally <40%) . Thanks to the development of instrumentation and chiral stationary phases (CSPs) in recent years, SFC is becoming the primary method for analytical and preparative separation of enantiomers in the pharmaceutical field .…”

Simultaneous determination of 17 bisphenols in polycarbonate by ultra‐high performance supercritical fluid chromatography with tandem mass spectrometry

“…When pressure goes up, the density of CO 2 increases, which will improve elution ability and shorten analysis time. Resolution was introduced to evaluate the influence of backpressure, column temperature and flow rate on chromatographic separation . At a backpressure of 11.72 MPa, peak 1/2, peak 6/7, and peak 10/11 have better resolution.…”

“…Ultra‐high performance supercritical fluid chromatography (UHPSFC) system has been introduced by Waters Corporation and attracted wide attention since 2012. Carbon dioxide (CO 2 ) as the main constituent of mobile phase, which has lower viscosity and higher mobility than organic reagents like methanol and acetonitrile in HPLC, theoretically improving analysis efficiency three to five times . Also, with less organic reagent consumption, UHPSFC is considered to be an environment‐friendly analysis method .…”