Application of High Speed Countercurrent Chromatography (HSCCC) to the Isolation of Kavalactones

Abstract

High speed countercurrent chromatography (HSCCC) was used to isolate the major kavalactones kavain, 7,8-dihydrokavain, methysticin, 7,8-dihydromethysticin, yangonin, and demethoxyyangonin. An ethanolic crude extract of kava root was subjected to HSCCC separation. Due to the presence of polymeric material, the sample load was limited. Hence, gel chromatography was applied in order to concentrate the kavalactones and to reduce the amount of polymers. The purified extract was again subjected to a HSCCC separation. A higher sample input was achieved, but only kavain and demethoxyyangonin were obtained in pure form. The remaining kavalactones eluted in three fractions, which needed to be further purified by preparative HPLC.

Performance Characteristics of Countercurrent Separation in Analysis of Natural Products of Agricultural Significance

“…The relative proportions of the constituent solvents within a family can be modified almost endlessly; therefore, organized systems of solvent system family members have been developed in the CS literature – . Biphasic solvent systems composed of varying concentrations of hexane−ethyl acetate−methanol–water are used extensively to separate and isolate phytochemicals from extracts – . The HEMWat family of 17 hexane−ethyl acetate−methanol–water solvent systems has been constructed with a progression of polarity from most polar (+8) to least polar (−8), as shown in Table .…”

Fast Isolation of Flavonoids from the Endemic Species Nolana ramosissima I.M. Johnst and Its Endothelium-Independent Relaxation Effect in Rat Aorta

“…Five major flavones were quickly isolated by HSCCC from a methanol extract of the endemic species N. ramosissima, and four of them showed relaxation activity. Besides, some 61 compounds were detected in both N. ramosissima polar extracts by UHPLC-MS. Of those, four were coumarins (peaks 2, 3, 6 and 18), 15 flavanones (peaks 4, 7, 9, 11, 15, 17, 21, 22, 32, 38, 46-49 and 56), 12 flavones (peaks 12, 13, 19, 23, 30, 39, 40-44 and 50), 3 phenolic acids (peaks 5,8 and 10) and 22 oxylipins/fatty acids (peaks 1,14,16,20,24,29,31,[33][34][35][36][37], 51-55, 57-61). Compound 2 presented higher relaxation effects than N. ramosissima extract.…”

“…This methodology was broadly used to separate the flavonoids from plants and fruits [10][11][12][13]. HSCCC offer important advantages in separation of natural products: lower consumption of solvents, use of green chemistry solvents, such as water and ethyl acetate, no absorption on solid surfaces such as conventional column chromatography, very higher amounts of processing sample, introduction of crude extracts, and full recovery of natural products [14][15][16][17][18]. In this work we have applied this technique for the fast detection of flavonoids, from the methanolic extract of N. ramosissima for the testing of their relaxation activity in rat aorta.…”

Countercurrent Separation of Natural Products

“…Schäfer and Winterhalter compared direct injection of crude kava root extract with injection of an extract that was fractionated on a Sephadex LH-20 column. In this case, prefractionation allowed for higher loading capacity and better resolution of target kava-lactones in the CS step . In almost a quarter of the surveyed articles, more than one SS was used to fractionate the target compounds.…”

“…(c) The recycling strategy. The same SS that was used in the first CS experiment may be employed again to recycle primary fractions by repeated CS . Han et al used a typical recycling arrangement to facilitate this operation .…”