Semi‐preparative isolation and purification of phenolic compounds from Achyrocline satureioides (Lam) D.C. by high‐performance counter‐current chromatography

Abstract

Introduction Phenolic compounds present in Achyrocline satureioides are known to have therapeutic benefits like antioxidant, anti‐inflammatory, and antitumour properties. The main polyphenols present in the plant are quercetin (QCT), luteolin (LUT), 3‐O‐methylquercetin (3OMQ), and achyrobichalcone (ACB). However, the effective isolation and purification of these compounds from A. satureioides inflorescences are not an easy task. Objective To develop an efficient high‐performance counter‐current chromatography (HPCCC) method for quick separation and purification of naturally occurring phenolic compounds from the extract of A. satureioides. Methodology A two‐step HPCCC semi‐preparative isolation method was developed using a solvent system composed of n‐hexane/ethyl acetate/methanol/water (0.8:1.0:0.8:1.0) and dichloromethane/methanol/water (3.5:3.5:2.5). Results The HPCCC method was used to obtain two fractions. The first fraction (F1) contained high levels of ACB, among other constituents, while the second fraction (F2) contained mostly QCT, LUT, and 3OMQ. Besides the high ACB content, F1 contained three other flavonoid‐aglycones (kaempferol, 97.3%; isokaempferide, 92.4%; and 3,3′‐di‐O‐methylquercetin, 95.2%) identified by an ultra‐performance liquid chromatography system coupled to a quadrupole time‐of‐flight with high‐definition mass spectrometry (UPLC‐QTOF/HDMS) and nuclear magnetic resonance (NMR) analysis. Purity levels of ACB, 3OMQ, QCT, and LUT were 98.0, 97.0, 97.5, and 90.2%, respectively. Conclusion This is the first time that high purity ACB and six other flavonoids were obtained from A. satureioides inflorescences by HPCCC. These excellent results reveal the potential and versatility of HPCCC as a technique to produce different types of products from this plant species on a semi‐preparative scale: enriched fractions, new metabolites, or high purity compounds.

Box-Behnken Design for Extraction Optimization Followed by High Performance Countercurrent Chromatography: Production of a Flavonoid-enriched Fraction from Achyrocline Satureioides

“…These biological properties have been very often ascribed to the high content of phenolic constituents in the inflorescences. Thus, the extraction and isolation of the flavonoids QCT, LUT, and 3OMQ has been the goal of many studies [2,9,10].…”

Achyrocline satureioides compounds, achyrobichalcone and 3‐O‐methylquercetin, induce mitochondrial dysfunction and apoptosis in human breast cancer cell lines

“…3OMQ is the most abundant among the flavonoids-aglycones in the cultivated species. 5 ACB is a novel molecule, reported only for A. satureioides, with a similar structure to other natural bichalcones. [6][7][8] 3OMQ possess biological properties and, in particular, has cytotoxic effects against the breast cancer cell lines SK-BR-3 and MCF-7, including drug-resistant strains.…”

“…The separation and isolation of ACB and 3OMQ from the dry extract was performed as previously described, using high-performance countercurrent chromatography (HPCCC, Dynamic Extraction). 5 The instrument was equipped with polytetrafluoroethylene (PTFE) bobbins with 1.6 mm inner diameter of tubing, 136 ml total volume and 6 ml sample loop. The solvent was pumped into the column with isocratic pumps (Scientific Systems, Inc., PA) at a flow rate of 6.0 ml/min while the apparatus was run at a rotation speed of 1,600 rpm.…”

Application of choline chloride deep eutectic solvents and high‐speed counter‐current chromatography to the extraction and purification of flavonoids from the thorns of Gleditsia sinensis Lam

“…Generally, a suitable composition for the twophase solvent system is crucial for a perfect HSCCC separation, which relied on satisfactory partition coefficient (K D ) values ranging from 0.5 to 2.0 for the target components. 25,26 Previous studies have identified the flavonoids from the thorns of G. sinensis mainly as flavonoid aglycones as well as a few flavonoid glycosides. Thus, several solvent systems of n-hexane-ethyl acetatemethanol-water, n-hexane-ethyl acetate-n-butanol-water, chloroform-methanol-water, dichloromethane-methanol-water, and dichloromethane-methanol-n-butanol-water with different composition ratios, were applied for testing the K D values of the five main constituents (1)(2)(3)(4)(5) in the DES extracts (Table 2).…”