I have ethyle acetate extracts of photorhbdus bacteria which need to be identified but I am confused, when we are running TLC or HPLC to detect secondary metabolites, then d'nt know what to use as the standard in it.
Thank you Best Regards
SUMAN
Why are you working on this project? That will determine what standard compound(s) to use. Do a literature search on your bacteria and see what compounds have already been produced. Chances are you will find one, or more, of those. Are you looking for antibiotic compounds, or the compounds that cause bioluminescence in that bacteria?
As for the solvent system, I assume silica TLC. As the extract came from ethyl acetate, I would start with silica with hexane/ethyl acetate, and silica with dichloromethane/methanol because ethyl acetate is medium polarity. For reverse phase, I might try water/acetonitrile. In either case, you will see a range of compounds.
Please look at the following below links which may help you in your analysis:
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Supelco/Brochure/1/tlc-brochure-mk.pdf
Thanks
Actually, I use TLC/HPLC for qualitative identification (separation) of crude extracts but dealing with secondary metabolites (natural products) need more purification to finally isolate pure compounds through column chromatography, then doing full characterization (NMR, FTIR, MS, elemental analysis and X-RAY crystallography). From the previously published articles, you can determine which classes of metabolites you deal with. Then if you don’t find the proper commercial standard available, you can use FTIR, GCMS, SDS-PAGE and TLC (RF and spot color) to confirm your compounds. Here are some articles may help you.
https://benthamopen.com/contents/pdf/TOTNJ/TOTNJ-3-83.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5487437/pdf/fmicb-08-01142.pdf
http://ibbj.org/article-1-110-fa.pdf
For standards.
https://www.analytics-shop.com/gb/chemicals/standards-and-reference-material/standards-according-to-application/environment.html
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