A marine bacterium demonstrates great promise in glycolipid production using waste frying soybean oil as substrate. Periodic determinations of residual oil in fermentation broth is important for studying the kinetics of production. It is necessary to determine the separate amounts of waste oil lipids (substrate) and glycolipid complex (product). Their concentrations are necessary for development of kinetic models for the fermentation process
Hello Maurice,
According to my experiences, different glycolipids have slightly different properties and require different separation strategies. Some glycolipids are more membrane-bond (e.g. trehalose lipids), some are not so membrane-bond (e.g. rhamnolipids). For membrane-bond glycolipids, sonication or exhaustive extraction may be needed.
When you have your glycolipids mixed with the substrate in the extraction solvent, the separation is mostly based on their differences in terms of polarity and affinity towards the column materials.
You have soybean oil which contains mostly fatty acids and thus tend to have higher polarity compared with glycolipids in general.
I would recommend the following strategy for the separation:
Load your redissolved crude extracts on to a silica gel column. Wash the column with solvents with increased polarity (e.g. hexane; n-hexane/chloroform/acetic acid
(20 : 80 : 0Æ5, v/v/v); chloroform/methanol (95 : 5, v/v); chloroform/methanol/water (85 : 15 : 2, v/v/v); chloroform/methanol/water (65 : 15 : 2, v/v/v) and methanol). Collect the eluates and detect the presence of sugar (glycolipid) with TLC (e.g. using Anthone agent).
Hope this will help you get your product :)
Hello Qinhong, thank you very much for the suggestion you gave for separation of glycolipids from soybean oil substrate. I have tried it and it worked like magic. Thanks a lot
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