Hi, I am getting fractions from my column which are UV inactive. using 10% sulphuric acid, they become visible. i want to separate those compounds. But staining is destructive for compounds. what method should I adpot for their isolation?
Hi Sania,
Did you try to visualize them by exposing the TLC plate to iodine vapor? Are the spots visible? if yes, you can use iodine vapor to monitor the isolation as it is not destructive. Good luck.
USE prep HPLC with an appropriate column and corona charged aerosol detector (CAD) to see the non-chromophoric compounds. Fraction collect the sample. When you know how many compounds you need to isolate than you can try TLC.
Use chromatotron or what is now called centrifuge, is a preparative, centrifugally accelerated, radial, thin-layer chromatography.
Ayorinde Victor Ogundele sir, I'm visulizing them using iodine chamber and they become visible. but can I place prep. TLC in iodine chamber? won't it change/destroy the structure and activity of my compounds?
Sania Izhar , It wont! Look for a big Iodine chamber that can accommodate your prep-TLC plate. Place it in the chamber, mark the spot of interest when it become visible, then leave the plate on the bench for iodine to evaporate then scrape the spot and desorb the compound from the silica gel using appropriate solvent.
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining