Hi,
I have started with organic solvent extraction of spent medium or cell free supernatant with butanol, but an oil globule-like substance is being formed in organic layer.
I want to know how to get rid of it.
Thank you.
The second organic phase is formed when not all extracted components are fully soluble in the solvent used. Try to use more lipophylic higher alcohol, e. g. octanol or decanol, if possible, instead of butanol or add it to butanol. I wish you good luck in your screening of proper solvent.
You have to work with different solvent or combination of them as suggested by Stefan. My only concern is also to pay attention on post separation recovery of your material of interest while choosing a different solvent. I assume heavy -ols may give you difficulty to get rid of from your purified material. All the best!
If the third phase is dispersed one possibility is to centrifuge the tube, say some 3 mi. at 3000 rpm, usually the phases become clear, and solid layer is visible at the interface.
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