Mass Directed Purification of Glucoraphanin from Broccoli Seeds using Preparative Scale Hydrophilic-Interaction Chromatography (HILIC)

Library Number:
APNT10072782
Part Number:
720002699en
Author(s):
Andrew Aubin
Source:
Waters
Content Type:
Application Notes
Content Subtype:
Application Notes
Related Products:
 
3100 Mass Detector
 
3100 Mass Detector
 
3100 Mass Detector
 
 

Broccoli seeds have been shown to be rich in glucoraphanin (20 to 50 mg/g) and are easy to obtain, making them a good choice for the isolation of glucoraphanin on a preparative scale. This application describes the mass-directed purification of glucoraphanin from broccoli seeds using the Waters AutoPurification System with the 3100 Mass Detector and an Atlantis Hydrophilic-Interaction Chromatography (HILIC) Silica Column.

  • Preparatory-scale Hydrophilic-Interaction Chromatography (HILIC) provided a useful separation technique for the isolation of the polar glucoraphanin. Higher throughput is possible due to reduced fraction dry down times and the opportunity to use higher flow rates.
  • An increase in purity from 53% to 95% was accomplished using mass-directed purification for the isolation of glucoraphanin from broccoli seeds.
  • A total of 139 mg of glucoraphanin was isolated in this example. This amount is consistent with published values.
  • Exact mass TOF-MS analysis of the purified fraction confirmed the presence of glucoraphanin.

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