Seven compounds from Hibiscus mutabilis L. leaves were first successfully
achieved by two-step high-speed counter-current chromatography with two-phase
solvent system composed of n-butanol-ethyl acetate-water (1:6:9, v/v/v) and
n-hexane-ethyl acetate-methanol-water (3:5:3:5, v/v/v/v/). The critical
experimental parameters of first-step separation were optimized with response
surface methodology as follows: flow rate was 1.1 mL/min, revolution speed
was 800 rpm and temperature was 30?C. Under the optimal conditions, around
5.0 mg of salicylic acid, 13.6 mg of rutin, 5.5 mg of genistein were obtained
in 100 mg crude sample. Then, 9.2 mg of potengriffioside A, 4.7 mg of
kaempferol 3-O-rutinoside, 3.0 mg of steppogenin and 2.5 mg of emodin were
obtained by second-step separation. The purities of the seven compounds
determined by UPLC were 96.2%, 93.8%, 95.4%, 94.3%, 98.0%, 94.1% and 90.8%,
respectively. Their chemical structures were identified by electron spray
ionization mass spectroscopy (ESI-MS) and 1H, 13C nuclear magnetic resonance
(NMR). Furthermore, compound steppogenin and genistein were first reported
from Hibiscus mutabilis L. The purification method was simple, efficient and
evaded tedious separation process.
Green Route for the Isolation and Purification of Hyrdoxytyrosol, Tyrosol, Oleacein and Oleocanthal from Extra Virgin Olive Oil
