食品科学

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半制备型高效液相色谱法分离刺葡萄花色苷单体

王维茜1,邓洁红1,*,刘永红2   

  1. 1.湖南农业大学食品科技学院,湖南 长沙 410128;2.湖南生物机电职业技术学院,湖南 长沙 410127
  • 出版日期:2016-09-25 发布日期:2016-09-26
  • 通讯作者: 邓洁红
  • 基金资助:

    国家自然科学基金面上项目(31271836);湖南省教育厅研究生科研创新课题(CX2015B264)

Preparation of Monomeric Anthocyanins from Vitis davidii Foex by Semi-Preparative HPLC

WANG Weiqian1, DENG Jiehong1,*, LIU Yonghong2   

  1. College of Food Science and Technology, Hunan Agricultural University, Changsha 410128, China;
    2. Hunan Biological and Electromechanical Polytechnic, Changsha 410127, China
  • Online:2016-09-25 Published:2016-09-26
  • Contact: DENG Jiehong

摘要:

经大孔吸附树脂HP-20纯化刺葡萄花色苷的粗提物后,以半制备型高效液相色谱法分离得到高纯度的刺葡萄花色苷单体。以XCharge C18柱(20 mm×250 mm,10 μm)制备柱,考察梯度洗脱条件、流动相流速、进样量对刺葡萄花色苷分离的影响,确定了最佳制备条件为甲醇-3%甲酸溶液流动相梯度洗脱、流速15 mL/min、进样量1.2 mL,实现了2 种主要花色苷单体的分离及制备。经超高效液相色谱-四极杆飞行时间质谱鉴定,2 种花色苷单体分别是锦葵素-3,5-O-双葡萄糖苷和锦葵素-3,5-O-双葡萄糖苷-香豆酰,产品纯度分别达到了99.54%和98.28%。方法具有简单易行、经济快速、易于放大等特点,适用于刺葡萄花色苷标准品的大规模制备。

关键词: 刺葡萄, 锦葵素, 半制备液相色谱, 超高效液相色谱-四极杆飞行时间质谱

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 MSE
method using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF MSE)
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 MSE

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