Hai, currently I am running with bacterial survival study in toxin containing Phosphate buffer saline(PBS) incubated for 1 week.. To observe the reduction of toxin(p-cresol) level, need to detect in TLC.. Is there any procedure available to separate these toxin from PBS saline...
Dear Vijaya,
The best extraction method will be as follows:
1-To the solution of p-cresol in PBS add 1n HCl until reaching an acidic pH of 1-2. The acidification is done to reduce the solubility of p-cresol in the aquesous solution. p-cresol is a phenol derivative which is more soluble in basic medium than acidic.
2-Extract 3 times with ether (volume of ether and aqueous solution is 1:1).
3-Combine the ether extracts.
4-Dry over MgSO4 anhydrous.
5-Evaporate until dryness. The residue is p-cresol.
Hoping this will be helpful,
Rafik
Dear Sir...
Excellent!!!! thank you so much for such a kind of Valuable information... Surely I will follow it sir... But this will be more suited for one of my exp., with 100microgram of p-cresol per 10ml (PBS), But in another study i'm going to fix with 2microgram, so how come this will suit. Let me check... Thanks a lot Sir...
Dear Vijaya,
For small quantities, you might use PLC or HPLC. The extraction procedure will be the same, but the purification will be using one of the above mentioned methods.
Rafik
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