Chemical Forums
Chemistry Forums for Students => Analytical Chemistry Forum => Topic started by: Mocook on August 19, 2013, 02:48:10 PM
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Hello all,
How can i calibrate what solvent system is efficient for my mixture, is TLC solvent system practical to use (not the same column tho)? Im attempting to isolate a halogenated porphyrin.
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I suppose you could try it, if you've used ordinary TLC plates, and your HPLC column is definitely a normal phase column, and you stick to an isocratic run (one eluent, not mixing a gradient over time,) then yes you might get a good starting point for a separation. You won't have to change too many things, and it will be a start. But if things are very different, say you're using a reverse phase column, then no, you can't use TLC developing solvent to run the HPLC. In that case, you can start with a quick literature, or even a Google search for the analyte, to see what sort of conditions you can start at. In fact, probably best to work with the second option in any case. I only mentioned the first option in case you were really stubborn. ;) What column do you have? And are you certain your HPLC detector can see your analyte?
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And do try a search on this forum -- the topic of HPLC of dervitized porphyrins did come up once or twice recently.
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For example, look at this Google result I found for an extremely perflorinated porphyrin. http://www.researchgate.net/publication/230817114_Synthesis_of_highly_functionalized_fluorinated_porphyrins/file/504635147ec6db2e03.pdf Its mostly about the synthesis, but they do mention the HPLC analysis. They used a
very short normal phase HPLC column, and they're eluent is simply dichloromethane. (Actually, a variety of non-polar eluents were used, based around dichloromethane, using fairly long columns)
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And here's a typical reverse-phase method: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1154376/?page=1