Preparation of CDF
1. Bacteria cultures were grown up in Minimal medium to
late log phase (OD600 = 2.0).
2. Equal volume of ethyl acetate (Optima Grade, Fisher
Scientific) was then added and incubated, with shaking at 200 rpm, for 1
3. The flasks were then allowed to sit for 1 h, or as
long as it takes to separate the aqueous phase from the ethyl acetate
phase. Should the layers fail to separate, additional ethyl acetate is
added and step 3 is repeated.
4. Remove the top layer containing the ethyl acetate
using a glass pipette, and store in glass container. The ethyl acetate
is removed by blow-drying, using the house air-line.
5. The concentrated extract, containing CDF, is then
stored at -20 C.
HPLC analyses of CDF