Abstract:

A semi-preparative high performance liquid chromatographic (HPLC) method using an XCharge C18 preparative
column (20 mm × 250 mm, 10 μm) was established for the preparation of anthocyanins monomers with high purity from
crude extracts of Vitis davidii Foex after purification with macroporous resin HP-20. The effects of experimental conditions
including mobile phase gradient elution conditions, flow rate and injection volume were investigated on the separation
efficiency of anthocyanins. The optimal chromatographic conditions were determined as follows: mobile phase, a mixture of
methanol and 3% formic acid water for gradient elution; flow rate, 15 mL/min; and injection volume, 1.2 mL, which allowed
the isolation and preparation of two major anthocyanins monomers in 18 min. The two anthocyanins were identified by MS^E
method using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF MS^E)
as malvidin-3,5-O- diglucoside and malvidin-3,5-O-diglucoside-coumary with purities of 99.54% and 98.28%, respectively.
The developed procedure was simple, effective, scalable and economic. It could be a promising alternative procedure for
large-scale preparation of standard materials of anthocyanins from Vitis davidii Foex.

Key words: Vitis davidii Foex, malvidin, semi-preparative HPLC, UPLC-QTOF MS^E