I have separated mycelia from 15 day old broth culture of Aspergillus ochraceus , then I have acidified the filtrate with HCL, than added some DEE (Di ethyl ether), then added equal amount of chloroform and separated the lower layer.
I am getting viscous liquid in lower layer as shown in photographs.
How can I remove that viscous liquid from lower layer?
The observed viscosity is may be due to lipids, just add n-hexane to this and vortex vigorously for 1 min and then centrifuge the samples at 4000-6000 rpm for 5 min, you will get two layers, the upper layer will be of n-hexane with all the lipids remove that and repeat the step. Ochratoxin A will be in lower layer.
Good Luck
By liquid-liquid extraction method, you will not get pure ochratoxin a, you have to purify it by preparative HPLC.
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