I'm analyzing the supernatants of bacteria broth cultures to find out some bioactive compounds. After I mix the same volume of supernatant and organic solvent, there is a jelly like middle layer. How I separate the organic layer only? Finally I want to make organic layer dry and use the residues for further studies (MIC).
Hi Sachithra,
This happens whenever you do a liquid-liquid partition to separate the organic layers and aqueous layer. Try to do partitioning with more (3 X 100 ml) organic solvent and finally pool the layers. It you are using chloroform then it will be easy for you to collect through a separating funnel (lower portion). For removing the gelly like middle layer leave the pooled layer in for around 8 hours and slowly collect the lower organic layer (if you are using chloroform). As in the case of ethyl acetate you may collect the lower water layer and remove the gelly like substance along with it.
I will recommend you use a separating funnel carefully observing all precautions so that you don't outrun other layer of the liquid. Just follow the suggestions of Preethidan DS ..
Thank you all for your answers....I tried the centrifugation. but it was not success. Then I separate both jelly like substance and organic layer together and allowed to drying. Then I realized it was mixture of water and organic solvent. Because after evaporation, I could see water again.
Does that jelly like layer has some interesting compounds and it just waste and we can discard it?
what if some of the components remained in the layer formed b/w the two layers. and does that layer has some compounds? or it was only waste and we can discard it?
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