Hi i will be performed reverse-phase HPLC using C-18 column for identification and quantification of my interest compounds, did i need to use RP-18 plate too for TLC? Thanks.
Yes and no. Yes if you want to maintain the same polarity (and solubility), like API separation and purification. No if you want to change the polarity and extract impurities (like Vitamin D analysis).
It is optional, RP TLC can show you if there are compounds present that are difficult to (or will not) elute off the C18 column. I like to run RP -TLC of a sample in increasing organic mixtures until I get to pure methanol or acetonitrile. If something remains at the plate origin in the pure organic phase it probably will not elute from the C18 HPLC column. An alternative sample preparation (like SPE) is then recommended.
Do you have specific method for your target compound by HPLC C-18, I mean from published journals? If yes maybe you can directly use HPLC without any TLC to optimize the separation. If it is not, if I mean you can not find any publications that described your compound, so I think you can first try using TLC as described by Bruce above for preliminary experiments due the low cost of TLC plate. Herewith I attach a link of TLC data base, maybe you can find published method for your compound, best regards
http://www.camag.com/en/tlc_hptlc/customer_magazine_cbs/ccbs_database.cfm?method=query
yes, to understand which one you isolate or identificate with HPLC, there is a rapport between TLC and HPLC and also to see the same order of compounds in both technics
TLC gives you the opportunity to see the spots color which in many cases gives an advantage for the identification of isolated molecules.
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