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