Look for articles from Prof Du Toit Loots. I did attached one that may be applicable in your situation
You can check the following article
Fomina, M., Charnock, J. M., Hillier, S., Alvarez, R. & Gadd, G. M. (2007). Fungal transformations of uranium oxides. Environ Microbiol 9, 1696–1710.
Authors perform organic acids analysis produced by fungi and contained in the agar
Dear Marli, Jai & Xabier
Thanks for your valuable suggestions !!!!!!!!!!!!!!!!!!!!
Dear Rashid
There is no one method can be used for extraction of all antibiotics, because the antibiotics are various compounds. You have to extract the broth with different solvents with increasing polarity (hexane, methylene chloride, ether, ethyl acetate), evaporate the solvent and test presence of antibiotics. To analyze and purify your antibiotic from the active extract requires use of several chromatography techniques such as TLC and HPLC.
To extract the organic acids from bacterial broth, the best method is by using anion exchanger. The acids are then eluted by HCl solution. The organic acid are preferred to be analyzed by HPLC using anion column, as GC will require drevatization.
Dear Rashid, Do u know if the antibiotic produced by your bacteria/fungi are volatiles or non-volatiles? if volatiles then GC-MS is a good choice for chemical analysis. for this Ethylacetate is good choice for extracting non-polar and medium polar compound from the culture broth.
For non-volatiles you will have choice of using iso-butanol, iso-propyl alcohol that could extract more polar compounds in hydrophobic solvent.
you can also use acid/base extraction method as described by Mr. Ahmed
Try ion-exchange chromatograhy using ion exchange resins and opH gradients For the antibiotics, I have no experience.
For the extraction of antibiotics, you may use different solvents to test which extractant gives you best extraction. Another option can be the use of XAD-4 resin, just pass your cell free liquid culture from it (200 ml from one column, (3 cm into 25-30 cm) and then get adsorbed antibiotics from the column by passing methanol, reduce its volume and check for antibiotics extraction. Different antibiotics are extracted best with different methods or extractants.
Good Luck
I observed some of the students using same method (XAD-4) for organic acids extraction from root exudates.
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