Abstract:

Polyamide chromatography and preparative high performance liquid chromatography (HPLC) were used for
the separation and preparation of two isomers of hyperoside from Abelmoschus manihot (L.) Medicus flowers. The results
indicated that the 50% ethanol eluate contained a mixture of hyperoside and its isomers after polyamide chromatography.
Then the freeze-dried mixture was separated by preparative HPLC on Calesil A-120 column (250 mm × 21.2 mm, 10 μm)
with acetonitrile-0.2% aqueous acetic acid (15:85, V/V) as the mobile phase by injecting 20.0 mg of the sample at a flow rate
of 25.0 mL/min; the column temperature was 30 ℃, and detection wavelength was 280 nm. Two isomers were achieved and
identified by MS, 1H and 13C NMR as hyperoside and isoquercitrin with purities of more than 99% and 97%, respectively,
and both were stereoisomer.

Key words: Abelmoschus manihot (L.) Medicus, polyamide chromatography, preparative high performance liquid chromatography, hyperoside, isoquercitrin